Cell Therapy

Part I: Technologies, Ethics & Regulations

by
Prof. K. K. Jain
MD, FRACS, FFPM

Jain PharmaBiotech
Basel, Switzerland

July 2012

A Jain PharmaBiotech Report

AUTHOR'S

BIOGRAPHY

Professor K. K. Jain is a neurologist/neurosurgeon by training and has been working in

the biotechnology/biopharmaceuticals industry for several years. He received graduate
training in both Europe and USA, has held academic positions in several countries, and is
a Fellow of the Faculty of Pharmaceutical Medicine of the Royal College of Physicians of
UK. Currently, he is a consultant at Jain PharmaBiotech.
Prof. Jain is the author of 440 publications including 21 books (4 as editor) and 50 special
reports, which have covered important areas in biotechnology, gene therapy and
biopharmaceuticals. He has also written a textbook of gene therapy which is the first book
on this subject to be translated into the Chinese language. A book on gene therapy
companies was published in 2000 by John Wiley & Sons and the 2011 version is included
in a special report on gene therapy published by Jain PharmaBiotech. Other reports
written by Prof. Jain have been published by PJB Publications, D & MD, Reuter's
Business Insight, and Decision Resources Inc. Cell Therapy is one of the several in-house
publications of Jain PharmaBiotech.
Prof. Jain's recent books include "Handbook of Nanomedicine" (Springer/Humana 2008;
Chinese edition, Peking University Press 2011; 2nd edition 2012), Textbook of
Personalized Medicine (Springer 2009), "Handbook of Biomarkers" (Springer 2010),
Handbook of Neuroprotection (Springer 2011), Applications of Biotechnology in
Cardiovascular Therapeutics (Springer, 2011), and Applications of Biotechnology in
Neurology (Springer, 2012, in preparation).

July 2012 (first edition published July 2001)

Copyright 2012 by
Jain PharmaBiotech
Blsiring 7
CH-4057 Basel
Switzerland
Tel & Fax:
Email:
Web site:

+4161-6924461
info@pharmabiotech.ch
http://pharmabiotech.ch/

All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted
in any form or by any means, electronic, mechanical, photocopying, or otherwise without the prior written
permission of the Publisher. This report may not be lent, resold or otherwise traded in any manner without the
consent of the Publisher. While all reasonable steps have been taken to ensure the accuracy of the information
presented, the Publisher cannot accept responsibility for inadvertent errors or omissions.

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TABLE

OF

CONTENTS

0. Executive Summary .............................................................................. 23

1. Introduction to Cell Therapy ................................................................. 27
Introduction....................................................................................................................... 27
Historical landmarks of cell therapy ................................................................................... 27
Interrelationship of cell therapy technologies ................................................................... 29
Cells and organ transplantation ......................................................................................... 29
Cells and protein/gene therapy ......................................................................................... 30
Cell therapy and regenerative medicine ............................................................................. 31
Cells therapy and tissue engineering ................................................................................. 31
Therapy based on cells involved in disease ........................................................................ 32
Advantages of therapeutic use of cells............................................................................... 32
Cell-based drug delivery .................................................................................................... 33
Cells as vehicles for gene delivery ................................................................................. 33
Red blood cells as vehicles for drug delivery ................................................................... 33
Advantages of cell-based drug delivery .......................................................................... 34
Limitations of cell-based drug delivery ........................................................................... 34

2. Cell Therapy Technologies .................................................................... 35

Introduction....................................................................................................................... 35
Cell types used for therapy ................................................................................................ 35
Sources of cells .................................................................................................................. 35
Xenografts ...................................................................................................................... 36
Cell lines ......................................................................................................................... 36
Immortalized cells ....................................................................................................... 36
Blood component therapy ................................................................................................. 36
Therapeutic apheresis .................................................................................................. 36
Leukoreduction ........................................................................................................... 37
Platelet therapy .......................................................................................................... 37
Basic technologies for cell therapy .................................................................................... 38
Cell culture...................................................................................................................... 38
Automated cell culture devices...................................................................................... 38
Cell culture for adoptive cell therapy.............................................................................. 39
Observation of stem cell growth and viability .................................................................. 39
Companies involved in cell culture ................................................................................. 39
Cell sorting...................................................................................................................... 41
Flow cytometry ........................................................................................................... 41
A dielectrophoretic system for cell separation ................................................................. 42
Adult stem cell sorting by identification of surface markers ............................................... 42
ALDESORTER system for isolation of stem cells ............................................................... 42
Dynabead technology for cell sorting ............................................................................. 42
Elutra Cell Separation System .................................................................................... 43
Molecular beacons for specific detection and isolation of stem cells.................................... 43
Multitarget magnetic activated cell sorter ....................................................................... 43
Nanocytometry ........................................................................................................... 44
Scepter cytometer .................................................................................................... 44
Companies supplying cell sorters .................................................................................. 44
Cell analysis .................................................................................................................... 45
Cell analyzers ............................................................................................................. 45
In vivo cell imaging ..................................................................................................... 46
Measuring cell density ................................................................................................. 46
Single-cell gene expression analysis ................................................................................... 46
Preservation of cells ......................................................................................................... 47
Innovations in cryopreservation .................................................................................... 48
Packaging of cells............................................................................................................. 48
Selective expansion of T cells for immunotherapy ................................................................ 48
Cloning and cell therapy ................................................................................................... 49
Techniques for cell manipulation ........................................................................................ 50
Cell-based drug discovery .................................................................................................. 50
Advantages and limitations of cell-based assays for drug discovery ........................................ 50
Advantages and limitations of cell-based toxicity screening .............................................. 51
Quality control of cells for drug discovery ....................................................................... 51
Companies involved in cell-based drug discovery ................................................................. 51
Drug delivery systems for cell therapy ............................................................................... 53
Intravenous delivery of stem cells ...................................................................................... 53
Intraarterial delivery of stem cells ...................................................................................... 53
Pharmacologically active microcarriers ................................................................................ 54

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Devices for delivery of cell therapy ..................................................................................... 54

Artificial cells ..................................................................................................................... 55
Applications of artificial cells .............................................................................................. 55
Cell encapsulation .............................................................................................................. 56
Diffusion capsule for cells .................................................................................................. 56
Encapsulated cell biodelivery ............................................................................................. 56
Therapeutic applications of encapsulated cells ..................................................................... 57
Nitric oxide delivery by encapsulated cells ........................................................................... 58
Implantation of microencapulated genetically modified cells .................................................. 59
Ferrofluid microcapsules for tracking with MRI ..................................................................... 59
Companies involved in encapsulated cell technology ............................................................. 59
Electroporation .................................................................................................................. 60
Gene therapy ..................................................................................................................... 61
Cell-mediated gene therapy .............................................................................................. 61
Fibroblasts ................................................................................................................. 61
Chondrocyte ............................................................................................................... 62
Skeletal muscle cells ................................................................................................... 62
Vascular smooth muscle cells ....................................................................................... 63
Keratinocytes ............................................................................................................. 63
Hepatocytes ............................................................................................................... 63
Lymphocytes .............................................................................................................. 64
Mammalian artificial chromosomes ................................................................................ 64
In vivo tracking of cells ..................................................................................................... 64
Molecular imaging for tracking cells .................................................................................... 64
MRI technologies for tracking cells ................................................................................ 65
Superparamagnetic iron oxide nanoparticles as MRI contrast agents ................................. 66
Visualization of gene expression in vivo by MRI............................................................... 66
Role of nanobiotechnology in development of cell therapy ................................................ 67
Cell transplantation for development of organs ................................................................. 67
Cells transplantation and tolerance ................................................................................... 68
Strategies to improve tolerance of transplanted cells ............................................................ 68
Encapsulation to prevent immune rejection .................................................................... 68
Prevention of rejection of xenotransplants ...................................................................... 68
Expansion of allospecific regulatory T cells ..................................................................... 69
Removal and replacement of pathogenic cells of the body ................................................. 69
Therapeutic leukocytapheresis ........................................................................................... 69

3. Stem Cells ............................................................................................. 71

Introduction....................................................................................................................... 71
Biology of stem cells .......................................................................................................... 72
Embryonic stem cells ........................................................................................................ 72
Growth and differentiation of ESCs ..................................................................................... 72
Mechanisms of differentiation of ESCs................................................................................. 73
Chemical regulation of stem cell differentiation ............................................................... 73
In vitro differentiation of hESCs .................................................................................... 73
SIRT1 regulation during stem cell differentiation ............................................................. 73
Regulation of stem cell self-renewal and differentiation ......................................................... 74
hESCs for reprogramming human somatic nuclei ............................................................. 74
Stem cells differentiation in the pituitary gland ............................................................... 74
Influence of microenvironment on ESCs .............................................................................. 75
Role of genes in differentiation of ESCs ............................................................................... 75
Global transcription in pluripotent ESCs ......................................................................... 75
Role of p53 tumor suppressor gene in stem cell differentiation .......................................... 76
Role of Pax3 gene in stem cell differentiation .................................................................. 76
Signaling pathways and ESC genes ............................................................................... 76
Epigenetics of hESCs ........................................................................................................ 77
Chromatin as gene regulator for ESC development .......................................................... 77
Mechanism of regulation of stem cells for regeneration of body tissues ................................... 78
Role of microenvironments in the regulation of stem cells ................................................ 78
Regulation and regeneration of intestinal stem cells ........................................................ 78
Parthenogenesis and human stem cells ............................................................................... 79
Uniparental ESCs ........................................................................................................ 80
Bone marrow stem cells .................................................................................................... 80
Hematopoietic stem cells ............................................................................................. 80
Role of HSCs in the immune system .............................................................................. 82
Derivation of HSCs from ESCs....................................................................................... 83
Mesenchymal stem cells ............................................................................................... 83
Multipotent adult progenitor cells .................................................................................. 84
Side population (SP) stem cells ..................................................................................... 85
Differentiation of adult stem cells ....................................................................................... 85
Growth and differentiation of HSCs..................................................................................... 86

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Signaling pathways in the growth and differentiation of HSCs ........................................... 86

Mathematical modeling of differentiation of HSCs ............................................................ 87
Role of prions in self renewal of HSCs ............................................................................ 87
Sources of stem cells ......................................................................................................... 88
Sources of of human embryonic stem cells .......................................................................... 88
Nuclear transfer to obtain hESCs ........................................................................................ 88
Direct derivation of hESCs from embryos without nuclear transfer .......................................... 89
Alternative methods of obtaining hESCs .............................................................................. 89
Establishing hESC lines without destruction of embryo ..................................................... 90
Altered nuclear transfer ............................................................................................... 91
Small embryonic-like stem cells .................................................................................... 91
Advantages and disadvantages of ESCs for transplantation ................................................... 91
Use of ESC cultures as an alternative source of tissue for transplantation ................................ 92
Spermatogonial stem cells ................................................................................................ 93
Amniotic fluid as a source of stem cells ............................................................................... 93
Amniotic fluid stem cells for tissue repair and regeneration .............................................. 94
Generation of iPS cells from AF cells .............................................................................. 94
Placenta as source of stem cells ......................................................................................... 95
Amnion-derived multipotent progenitor cells ................................................................... 95
Placenta as a source of HSCs ........................................................................................ 95
Umbilical cord as a source of MSCs ................................................................................ 96
Umbilical cord blood as source of neonatal stem cells ........................................................... 96
Cryopreservation of UCB stem cells ............................................................................... 97
UCB as source of MSCs ................................................................................................ 97
Applications of UCB ..................................................................................................... 97
Advantages of UCB...................................................................................................... 98
Limitations of the use of UCB ........................................................................................ 98
Licensing and patent disputes involving UCB .................................................................. 99
Infections following UCB transplants ............................................................................ 100
Unanswered questions about UCB transplantation ......................................................... 100
Companies involved in UCB banking ............................................................................ 101
UCB banking in the UK............................................................................................... 102
US national UCB banking system ................................................................................ 102
Future prospects of UCB as a source of stem cells ......................................................... 103
Induced pluripotent stem cells derived from human somatic cells ................................... 104
Characteristics of iPSCs .................................................................................................. 104
DNA methylation patterns of iPS cells ............................................................................... 105
iPSCs derived from oocytes through SCNT......................................................................... 105
iPSCs derived from skin .................................................................................................. 105
iPSCs derived from blood ................................................................................................ 106
Use of retroviral vectors for generation of iPSCs ................................................................ 107
Use of non-integrating viral vectors for generation of iPSCs ................................................. 107
Generation of clinically relevant iPSCs .............................................................................. 108
Generation of RBCs from iPSCs ................................................................................... 108
iPSCs and disease modeling ....................................................................................... 109
iPSCs for patient-specific regenerative medicine............................................................ 109
Concluding remarks about clinical potential of iPSCs ...................................................... 110
Induced conditional self-renewing progenitor cells ......................................................... 110
Epiblast stem cells ........................................................................................................... 111
Comparison of development of human and mouse ESCs ..................................................... 111
Conversion of hESCs to mouse ESC-like nave states .......................................................... 111
Sources of adult human stem cells................................................................................... 112
Adipose tissue as a source of stem cells ............................................................................ 112
Intravenous infusion of adipose tissue derived MSCs ..................................................... 112
iPSCs derived from adult human adipose stem cells ....................................................... 113
Regulation of adipose stem cells differentiation ............................................................. 113
Transforming adult adipose stem cells into other cells.................................................... 113
Multipotent stem-like cells derived from vascular endothelial cells ........................................ 114
Skin as a source of stem cells .......................................................................................... 114
Controlling the maturation of embryonic skin stem cells ................................................. 114
Epidermal neural crest stem cells ................................................................................ 114
Follicle stem cells ...................................................................................................... 115
Mesenchymal stem cells in skin................................................................................... 115
Regulation of stem cells in hair follicles ........................................................................ 116
Skin-derived precursor cells ....................................................................................... 116
Stem cells in teeth ......................................................................................................... 116
Peripheral blood stem cells .............................................................................................. 117
Spleen as a source of adult stem cells .............................................................................. 117
Search for master stem cells ........................................................................................... 118
Vascular cell platform to self-renew adult HSC ................................................................... 118
Adult stem cells vs embryonic stem cells ......................................................................... 119

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Biological differences between adult and embryonic stem cells ............................................ 119
Neural crest stem cells from adult hair follicles .................................................................. 119
Transdifferentiation potential of adult stem cells ................................................................ 119
Limitations of adult stem cells ......................................................................................... 121
Comparison of human stem cells according to derivation ................................................ 121
VENT cells ........................................................................................................................ 121
Stem cell banking ............................................................................................................ 122
Stem cell technologies ..................................................................................................... 122
Analysis of stem cell growth and differentiation ................................................................. 122
Activation of bone marrow stem cells into therapeutic cells ................................................. 123
Role of nitric oxide in stem cell mobilization and differentiation ............................................ 123
Role of natriuretic peptide receptor-C in self-renewal of murine ESCs ................................... 123
Tracking self-renewal and expansion of transplanted muscle stem cells ................................ 124
Stem cell biomarkers ...................................................................................................... 124
Endoglin as a functional biomarker of HSCs .................................................................. 124
STEMPRO EZChek for analysis of biomarkers of hESCs .............................................. 125
SSEA-4 as biomarker of MSCs .................................................................................... 125
p75NTR as a biomarker to isolate adipose tissue-derived stem cells ................................ 125
Neural stem cell biomarker ......................................................................................... 125
Protein expression profile as biomarker of stem cells ..................................................... 126
Real-time PCR for quantification of protein biomarkers................................................... 126
Study of stem cell pathways ............................................................................................ 126
Stem cell genomics ........................................................................................................ 127
Gene expression in hESCs .......................................................................................... 127
Genomic alterations in cultured hESCs ......................................................................... 127
Study of transcriptional regulation of stem cell genes .................................................... 128
Casanova gene in zebrafish ........................................................................................ 128
Nanog gene .............................................................................................................. 128
Gene inactivation to study hESCs ..................................................................................... 130
RNAi to study gene inactivation in hESCs ..................................................................... 130
Study of ESC development by inducible RNAi................................................................ 130
Targeting Induced Local Lesions in Genomes ................................................................ 131
Homologous recombination of ESCs ............................................................................. 131
Immortalization of hESCs by telomerase ...................................................................... 131
Gene modification in genomes of hESCs and hiPSCs using zinc-finger nuclease ...................... 132
miRNA and stem cells ..................................................................................................... 132
Role of miRNAs in gene regulation during stem cell differentiation ................................... 132
Influence of miRNA on stem cell formation and maintenance .......................................... 133
Transcriptional regulators of ESCs control miRNA gene expression .................................. 134
Stem cells and cloning .................................................................................................... 134
Cell nuclear replacement and cloning ........................................................................... 134
Nuclear transfer and ESCs .......................................................................................... 134
Cloning from differentiated cells .................................................................................. 136
Cloning mice from adult stem cells .............................................................................. 136
Creating interspecies stem cells .................................................................................. 137
Cloned cells for transplantation medicine ..................................................................... 137
Claims of cloning of hESCs ......................................................................................... 137
Cytogenetics of embryonic stem cells ............................................................................... 139
Stem cell proteomics ...................................................................................................... 139
Comparative proteomic analysis of somatic cells, iPSCs and ESCs ................................... 140
hESC phosphoproteome ............................................................................................. 140
Proteomic studies of mesenchymal stem cells ............................................................... 140
Proteomic profiling of neural stem cells ........................................................................ 141
Proteome Biology of Stem Cells Initiative ..................................................................... 141
Technologies for mobilization, expansion, and engraftment of stem cells............................... 141
Chemoattraction of neuronal stem cells through GABA receptor ...................................... 142
Enhancement of HSC engraftment by calcium-sensing receptor ...................................... 142
Ex vivo expansion of human HSCs in culture ................................................................ 143
Ex vivo expansion of MSCs ......................................................................................... 143
Ex vivo expansion of UCB cells for transplantation ......................................................... 144
Expansion of adult stem cells by activation of Oct4 ........................................................ 144
Expansion of transduced HSCs in vivo.......................................................................... 144
Expansion of stem cells in vivo by Notch receptor ligands............................................... 144
In vivo adipogenesis induced by adipose tissue-derived stem cells .................................. 145
Selective mobilization of progenitor cells from bone marrow ........................................... 145
Selective Amplification ............................................................................................... 145
Synthetic substrates for ESC growth and expansion ...................................................... 146
Technologies for inducing differentiation of stem cells ......................................................... 146
Enhancement of stem cell differentiation by Homspera .................................................. 146
Generation of RBCs from HSCs ................................................................................... 146
Generation of multiple types of WBCs from hESCs and iPSCs .......................................... 147

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Growth factor-induced differentiation of MAPCs ............................................................. 147

Lineage selection to induce differentiation of hESCs ....................................................... 147
Mechanical strain to induce MSC differentiation ............................................................. 148
Neurotrophin-mediated survival and differentiation of hESCs .......................................... 148
Synthetic biology and stem cells ................................................................................. 149
Use of RNAi to expand the plasticity of autologous adult stem cells ................................. 149
Use of carbohydrate molecules to induce differentiation of stem cells .............................. 149
Limitations of the currently available stem cell lines in the US ............................................. 149
Stem cell separation ....................................................................................................... 150
Stem cell culture ............................................................................................................ 150
Culture of hMSCs ...................................................................................................... 151
Elimination of contaminating material in stem cell culture .............................................. 151
Long-term maintenance of MSC multipotency in culture ................................................. 153
Nanofiber scaffolds for stem cell culture ....................................................................... 153
Conversion of stem cells to functioning adipocytes ............................................................. 154
Mass production of ESCs ................................................................................................. 154
Promoting survival of dissociated hESCs ........................................................................... 154
Analysis and characterization of stem cells ........................................................................ 155
Havesting and identification of EPCs ................................................................................. 155
Labeling of stem cells ..................................................................................................... 155
Labeling, imaging and tracking of stem cells in vivo ........................................................... 156
Perfluorocarbon nanoparticles to track therapeutic cells in vivo ....................................... 156
Project for imaging in stem cell therapy research .......................................................... 156
Quantum dots for labeling and imaging of stem cells ..................................................... 157
Superparamagnetic iron oxide nanoparticles for tracking MSCs ....................................... 157
Applications of stem cells ................................................................................................ 157
Commercial development and applications of adult stem cells .............................................. 158
Retrodifferentiation of stem cells ................................................................................. 158
MultiStem ................................................................................................................ 158
Controlling the maintenance process of hematopoietic stem cells ......................................... 159
Self renewal and proliferation of HSCs ......................................................................... 159
Aging and rejuvenation of HSCs .................................................................................. 159
Peripheral blood stem cell transplantation ......................................................................... 160
Role of stem cells in regeneration .................................................................................... 160
Promotion of regeneration by Wnt/beta-catenin signaling ............................................... 160
Stem cell activation for regeneration by using glucocortoids ........................................... 161
Stem cells and human reproduction ................................................................................. 161
Expansion of spermatogonial stem cells ....................................................................... 161
Conversion of ESCs into spermatogonial stem cells ....................................................... 161
Conversion of stem cells to oocytes ............................................................................. 162
ESCs for treatment of infertility in women .................................................................... 162
Cloning human embryos from oocytes matured in the laboratory .................................... 163
In utero stem cell transplantation................................................................................ 163
Innovations in delivery of stem cells ................................................................................. 164
Polymeric capsules for stem cell delivery ...................................................................... 165
Immunological aspects of hESC transplantation ................................................................. 165
Immunosuppression to prevent rejection of hESC transplants ......................................... 165
Histocompatibility of hESCs ........................................................................................ 165
Strategies for promoting immune tolerance of hESCs .................................................... 166
Stem cells for organ vascularization ................................................................................. 166
Activation of EphB4 to enhance angiogenesis by EPCs ................................................... 167
Advantages and limitations of clinical applications of MSCs .................................................. 167
Biofusion by genetically engineering stem cells .................................................................. 168
Stem cell gene therapy ................................................................................................... 168
Combination of gene therapy with nuclear transfer ........................................................ 168
Gene delivery to stem cells by artificial chromosome expression ..................................... 169
Genetic manipulation of ESCs ..................................................................................... 169
Genetic engineering of human stem cells for enhancing angiogenesis .............................. 169
HSCs for gene therapy ............................................................................................... 170
iPSCs for targeted gene correction of 1-antitrypsin deficiency ....................................... 171
Helper-dependent adenoviral vectors for gene transfer in ESCs....................................... 171
Lentiviral vectors for in vivo gene transfer to stem cells ................................................. 171
Linker based sperm-mediated gene transfer technology ................................................. 171
Mesenchymal stem cells for gene therapy .................................................................... 172
Microporation for transfection of MSCs ......................................................................... 172
Regulation of gene expression for SC-based gene therapy.............................................. 172
Stem cells and in utero gene therapy........................................................................... 173
Therapeutic applications for hematopoietic stem cell gene transfer .................................. 173
The future of hematopoietic stem cell gene therapy....................................................... 173
Stem cell pharmaceutics .................................................................................................. 173
Pharmaceutical manipulation of stem cells ........................................................................ 174

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Antisense approach for preservation and expansion of stem cells .................................... 175
Expansion of HSCs in culture by inhibiting aldehyde dehydrogenase ................................ 175
Manipulation of stem cells with growth factors .............................................................. 176
Mobilization of stem cells by cytokines/chemokines ....................................................... 178
Mobilization of adult human HSCs by use of inhibitors.................................................... 179
Mobilization of stem cells by HYC750 ........................................................................... 179
Mobilization of stem cells by hyperbaric oxygen ............................................................ 180
Mobilization by adenoviral vectors expressing angiogenic factors..................................... 180
Stem cell mobilization by acetylcholine receptor agonists ............................................... 180
Use of parathyroid hormone to increase HSC mobilization .............................................. 181
Use of small molecule compounds for expansion of HSCs ............................................... 181
Role of stem cells in therapeutic effects of drugs ................................................................ 181
Stem cells for drug discovery........................................................................................... 182
Target identification .................................................................................................. 182
High-throughput screening ......................................................................................... 182
Cardiomyocytes derived from hESCs ........................................................................... 182
ESCs as source of models for drug discovery ................................................................ 183
hESC-derived hepatocytes for drug discovery ............................................................... 184
Advantages and limitations of use of stem cells for drug discovery .................................. 184
Stem cells for drug delivery ............................................................................................. 185
Toxicology and drug safety studies using ESCs versus other cells ......................................... 185
Future challenges for stem cell technologies ................................................................... 188
Study of the molecular mechanism of cell differentiation ..................................................... 188
MBD3-deficient ESC line.................................................................................................. 188
In vivo study of human hemopoietic stem cells .................................................................. 189
Stem cell biology and cancer ........................................................................................... 189
Research into plasticity of stem cells from adults ............................................................... 190
Stem cells and aging ...................................................................................................... 190
Hybrid embryos/cybrids for stem cell research ................................................................... 191
Generation of patient-specific pluripotent stem cells ........................................................... 192
Markers for characterizing hESC lines ............................................................................... 192
Switch of stem-cell function from activators to repressors ................................................... 193
Stem cell research at academic centers ........................................................................... 193
International Regulome Consortium ................................................................................. 194
Companies involved in stem cell technologies ................................................................. 195
Concluding remarks about stem cells............................................................................... 200
Challenges and future prospects of stem cell research .................................................... 200

4. Clinical Applications of Cell Therapy ................................................... 203

Introduction..................................................................................................................... 203
Cell therapy for hematological disorders.......................................................................... 203
Transplantation of autologous hematopoietic stem cells ...................................................... 203
Hemophilias .................................................................................................................. 203
Ex vivo cell/gene therapy of hemophilia B .................................................................... 203
Cell/gene therapy of hemophilia A ............................................................................... 204
Hematopoietic stem cell therapy for thrombocytopenia ....................................................... 205
Stem cell transplant for sickle cell anemia ......................................................................... 205
Treatment of chronic acquired anemias............................................................................. 206
Implantation of genetically engineered HSCs to deliver rhEpo ......................................... 206
Drugs acting on stem cells for treatment of anemia ....................................................... 206
Stem cell therapy of hemoglobinopathies .......................................................................... 207
Stem cells for treatment of immunoglobulin-light chain amyloidosis ..................................... 207
Future prospects of cell therapy of hematological disorders ................................................. 207
Cell therapy for immunological disorders ......................................................................... 208
Role of dendritic cells in the immune system ..................................................................... 208
Modifying immune responses of DCs by vaccination with lipiodol-siRNA mixtures ................... 208
Potential of MSCs as therapy for immune-mediated diseases ............................................... 209
Stem cell therapy of chronic granulomatous disease ........................................................... 209
Stem cell therapy of X-linked severe combined immunodeficiency ........................................ 210
Stem cell therapy of autoimmune disorders ....................................................................... 210
Treatment of rheumatoid arthritis with stem cells.......................................................... 210
Treatment of Crohn's disease with stem cells ................................................................ 211
Stem cell transplants for scleroderma .......................................................................... 211
Role of T Cells in immunological disorders ......................................................................... 212
Autologous T cells from adult stem cells ....................................................................... 212
Cell therapy for graft vs host disease ................................................................................ 213
MSCs for GVHD ......................................................................................................... 214
Cell therapy for viral infections ........................................................................................ 214
T-cell therapy for CMV ............................................................................................... 214
T-cell therapy for HIV infection ................................................................................... 215
T-cell immunity by Overlapping Peptide-pulsed Autologous Cells ..................................... 215

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Anti-HIV ribozyme delivered in hematopoietic progenitor cells ........................................ 216

Dendritic-cell targeted DNA vaccine for HIV .................................................................. 216
Cell therapy of lysosomal storage diseases ...................................................................... 216
Niemann-Pick disease ..................................................................................................... 217
Gauchers disease .......................................................................................................... 217
Fabry's disease .............................................................................................................. 218
Cell therapy for diabetes mellitus .................................................................................... 218
Limitations of current treatment ...................................................................................... 219
Limitations of insulin therapy for diabetes mellitus ........................................................ 219
Limitations of pancreatic transplantation ...................................................................... 219
Islet cell transplantation ................................................................................................. 220
Autologous pancreatic islet cell transplantation in chronic pancreatitis ............................. 220
Clinical trials of pancreatic islet cell transplants for diabetes ........................................... 220
Drawbacks of islet cell therapy.................................................................................... 221
Use of an antioxidant peptide to improve islet cell transplantation ................................... 222
Cdk-6 and cyclin D1 enhance human beta cell replication and function ............................ 222
A device for delivery of therapeutic cells in diabetes ...................................................... 222
Monitoring of islet cell transplants with MRI .................................................................. 222
Concluding remarks about allogeneic islet transplantation for diabetes ............................ 223
Encapsulation of insulin producing cells............................................................................. 223
Encapsulated porcine pancreatic islet cells for pancreas ................................................. 223
Encapsulated insulinoma cells ..................................................................................... 224
Magnetocapsule enables imaging/tracking of islet cell transplants ................................... 224
Islet precursor cells ........................................................................................................ 225
Dedifferentiation of cells to promote regeneration ........................................................... 225
Pharmacological approaches for cell regeneration ............................................................ 226
Xenotransplantation of embryonic pancreatic tissue ........................................................... 226
Non-pancreatic tissues for generation of insulin-producing cells ........................................... 227
Exploiting maternal microchimerism to treat diabetes in the child ........................................ 227
Bio-artificial substitutes for pancreas ................................................................................ 227
Role of stem cells in the treatment of diabetes .................................................................. 228
Embryonic stem cells for diabetes ............................................................................... 228
HSC transplantation to supplement immunosuppressant therapy .................................... 229
Human neural progenitor cells converted into insulin-producing cells ............................... 229
Insulin-producing cells derived from UCB stem cells ...................................................... 230
iPS cells for diabetes ................................................................................................. 230
Isolation of islet progenitor cells.................................................................................. 230
Pancreatic progenitor cells Expansion in vitro ............................................................... 231
Pancreatic stem cells ................................................................................................. 231
Stem cell injection into portal vein of diabetic patients ................................................... 231
Dendritic cell-based therapy for type 1 diabetes................................................................. 232
Vaccine for diabetes ....................................................................................................... 232
Gene therapy in diabetes ................................................................................................ 232
Viral vectors for gene therapy of diabetes .................................................................... 233
Genetically engineered dendritic cells .......................................................................... 233
Genetically altered liver cells ...................................................................................... 233
Genetically modified stem cells ................................................................................... 234
Companies developing cell therapy for diabetes ................................................................. 234
Concluding remarks about cell and gene therapy of diabetes ............................................... 235
Cell therapy of gastrointestinal disorders ........................................................................ 236
Inflammatory bowel disease ............................................................................................ 236
Cell therapy for liver disorders......................................................................................... 237
Types of cells and methods of delivery for hepatic disorders ................................................ 237
Bioartificial liver ............................................................................................................. 238
Hepatocyte-based artificial liver .................................................................................. 238
Extracorporeal Liver Assist Device ............................................................................... 238
Limitations of bioartificial liver .................................................................................... 239
Proliferating cell-based bioartificial liver ....................................................................... 239
Stem cells for hepatic disorders ....................................................................................... 239
Deriving hepatocytes from commercially available hMSCs .............................................. 240
Implantation of hepatic cells derived from hMSCs of adipose tissue ................................. 240
Heterologous adult liver progenitor cells ...................................................................... 241
MSC derived molecules for reversing hepatic failure ...................................................... 241
Cell-based gene therapy for liver disorders ........................................................................ 241
Transplantation of genetically modified fibroblasts ......................................................... 242
Transplantation of genetically modified hepatocytes ...................................................... 242
Intraperitoneal hepatocyte transplantation ................................................................... 242
Genetically modified hematopoietic stem cells .............................................................. 242
Use of iPSCs derived from somatic cells for liver regeneration ......................................... 243
Clinical applications ........................................................................................................ 243
Future prospects of cell-based therapy of hepatic disorders ................................................. 243

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Cell therapy of renal disorders ......................................................................................... 244

Bioartificial kidney .......................................................................................................... 244
Cell-based repair for vascular access failure in renal disease ............................................... 245
Mesangial cell therapy for glomerular disease .................................................................... 245
Stem cells for renal disease ............................................................................................. 245
Role of stem cells in renal repair ................................................................................. 246
Bone marrow stem cells for renal disease ..................................................................... 246
MSC therapy for renal disease .................................................................................... 246
Cell therapy for pulmonary disorders ............................................................................... 247
Delivery of cell therapy for pumonary disorders ................................................................. 247
Intratracheal injection of cells for pulmonary hypoplasia ................................................ 247
Role of stem cells in pulmonary disorders.......................................................................... 247
Lung stem cells ......................................................................................................... 247
Lung tissue regeneration from stem cells ..................................................................... 248
Role of stem cells in construction of the Cyberlung ........................................................ 248
Respiratory epithelial cells derived from UCB stem cells ................................................. 248
Respiratory epithelial cells derived from hESCs ............................................................. 249
Lung tissue engineering with adipose stromal cells ........................................................ 249
Cell-based tissue-engineering of airway ....................................................................... 250
Pulmonary disorders that can be treatable with stem cells................................................... 250
Acute lung injury and ARDS treated with MSCs ............................................................. 251
Bronchopulmonary dysplasia treated with MSCs ............................................................ 251
Chronic obstructive pulmonary disease treated with MSCs .............................................. 252
Cystic fibrosis treatment with genetically engineered MSCs ............................................ 252
Lung regeneration by integrin 64-expressing alveolar epithelial cell ............................. 252
Pulmonary arterial hypertension treatment with EPCs .................................................... 253
Cell therapy for disorders of bones and joints .................................................................. 253
Repair of fractures and bone defects ................................................................................ 253
Adult stem cells for bone grafting ................................................................................ 254
Cell therapy for osteonecrosis ..................................................................................... 255
Cell therapy for cervical vertebral interbody fusion ........................................................ 255
Cell-mediated gene therapy for bone regeneration ........................................................ 255
ESCs for bone repair.................................................................................................. 255
Intrauterine use of MSCs for osteogenesis imperfecta .................................................... 256
In vivo bone engineering as an alternative to cell transplantation .................................... 256
MSCs for repair of bone defects .................................................................................. 256
MSCs for repair of bone fractures ................................................................................ 259
Osteocel .................................................................................................................. 259
Stem cells for repairing skull defects ........................................................................... 260
Stem cell-based bone tissue engineering ...................................................................... 260
Spinal fusion using stem cell-based bone grafts ............................................................ 261
Osteoarthritis and other injuries to the joints ..................................................................... 261
Mosaicplasty ............................................................................................................. 262
Autologous cultured chondrocytes ............................................................................... 262
Autologous intervertebral disc chondrocyte transplantation ............................................ 263
Cartilage repair by genetically modified fibroblasts expressing TGF- ............................... 264
Generation of cartilage from stem cells ........................................................................ 264
Repair of osteonecrosis by bone marrow derived MSCs .................................................. 266
Role of cell therapy in repair of knee cartilage injuries ........................................................ 266
Role of cells in the repair of anterior cruciate ligament injury .......................................... 268
Autologous tenocyte implantation in rotator cuff injury repair .............................................. 268
Platelet injection for tennis elbow ..................................................................................... 269
Cell therapy of rheumatoid arthritis .................................................................................. 269
Cell therapy for diseases of the eye ................................................................................. 270
Cell therapy for corneal repair ......................................................................................... 270
Stem cell therapy for limbal stem cell deficiency ................................................................ 271
Role of stem cells in fibrosis following eye injury ................................................................ 272
Stem cell transplantation for radiation sickness .............................................................. 272
MSCs for treatment of radiation damage to the bone .......................................................... 272
MSCs for regeneration of ovaries following radiotherapy damage ......................................... 273
Cell therapy for wound healing ........................................................................................ 273
Cells to form skin substitutes for healing ulcers ................................................................. 273
CellSpray for wound repair .............................................................................................. 274
Cell therapy for burns ..................................................................................................... 275
Closure of incisions with laser guns and cells ..................................................................... 276
Genetically engineered keratinocytes for wound repair........................................................ 276
Stem cells for skin regeneration ....................................................................................... 276
Follicular stem cells for skin and wound repair .............................................................. 276
MSCs for wound healing ............................................................................................. 277
Regeneration of aging skin by adipose-derived stem cells .............................................. 277
Reprogramming autologous stem cells for wound regeneration ....................................... 277

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Role of amniotic fluid MSCs in repair of fetal wounds ..................................................... 278

Concluding remarks on regeneration of skin by stem cells .............................................. 278
Cell therapy for regeneration ........................................................................................... 278
Stem cells for regenerating organs ................................................................................... 278
Umbilical cord blood for regeneration ............................................................................... 279
Role of stem cells in regeneration of esophageal epithelium ................................................ 279
Cell therapy for regeneration of muscle wasting ................................................................. 280
Role of cells in tissue engineering and reconstructive surgery ........................................ 280
Scaffolds for tissue engineering ....................................................................................... 280
Improving vascularization of engineered tissues ................................................................ 281
Repair of aging skin by injecting autologous fibroblasts....................................................... 281
Enhancing vascularization by combining cell and gene therapy ............................................ 282
Nanobiotechnology applied to cells for tissue engineering.................................................... 282
Choosing cells for tissue engineering ................................................................................ 283
Stem cells for tissue repair .............................................................................................. 283
ESCs vs adult SCs for tissue engineering ...................................................................... 283
Use of adult MSCs for tissue engineering ...................................................................... 284
Stem cells for tissue engineering of various organs ............................................................ 285
Breast reconstruction by adipose tissue-derived stem cells ............................................. 285
Engineering of healthy living teeth from stem cells ........................................................ 285
Intra-uterine repair of congenital defects using amniotic fluid MSCs ................................ 286
Skin regeneration by stem cells as an alternative to face transplant ................................ 287
Tissue engineering of bone by stem cells...................................................................... 287
Cell-based tissue engineering in genitourinary system ........................................................ 287
Urinary incontinence .................................................................................................. 287
Tissue engineering of urinary bladder .......................................................................... 289
Label retaining urothelial cells for bladder repair ........................................................... 289
MSCs for bladder repair ............................................................................................. 289
Tissue-engineering of urethra using autologous cells ..................................................... 290
Repair of the pelvic floor with stem cells from the uterus ............................................... 290
Reconstruction of vagina from stem cells ..................................................................... 290
Reconstruction of cartilage for repair of craniofacial defects ................................................. 291
Cell therapy for rejuvenation ........................................................................................... 291
Cell therapy for performance enhancement in sports ...................................................... 291
Application of stem cells in veterinary medicine .............................................................. 292
Use of stem cells to repair tendon injuries ......................................................................... 292
Stem cells for spinal cord injury in dogs ............................................................................ 292

5. Cell Therapy for Cardiovascular Disorders .......................................... 295

Introduction to cardiovascular disorders ......................................................................... 295
Limitations of current therapies for myocardial ischemic disease .................................... 295
Types of cell therapy for cardiovascular disorders ........................................................... 295
Cell-mediated immune modulation for chronic heart disease................................................ 296
Human cardiovascular progenitor cells .............................................................................. 297
Inducing the proliferation of cardiomyocytes ..................................................................... 297
Pericardial origin of colony-forming units .......................................................................... 298
Role of the SDF-1-CXCR4 axis in stem cell therapies for myocardial ischemia ........................ 298
Role of splenic myocytes in repair of the injured heart ........................................................ 298
Reprogramming of fibroblasts into functional cardiomyocytes .............................................. 299
Small molecules to enhance myocardial repair by stem cells................................................ 299
Cell therapy for atherosclerotic coronary artery disease .................................................. 299
MyoCell (Bioheart)....................................................................................................... 300
Cardiac stem cells .......................................................................................................... 300
Cardiomyocytes derived from epicardium .......................................................................... 301
Methods of delivery of cells to the heart .......................................................................... 302
Cellular cardiomyoplasty ................................................................................................. 302
IGF-1 delivery by nanofibers to improve cell therapy for MI ................................................. 302
Non-invasive delivery of cells to the heart by Morphguide catheter .................................... 302
Cell therapy for cardiac revascularization ........................................................................ 303
Transplantation of cardiac progenitor cells for revascularization of myocardium ..................... 303
Stem cells to prevent restenosis after coronary angioplasty ................................................. 303
Role of cells in cardiac tissue repair ................................................................................. 304
Modulation of cardiac macrophages for repair of infarct ...................................................... 304
Transplantation of myoblasts for myocardial infarction ........................................................ 304
Patching myocardial infarction with fibroblast culture.......................................................... 305
Cardiac repair with myoendothelial cells from skeletal muscle.............................................. 305
Myocardial tissue engineering .......................................................................................... 306
Role of stem cells in repair of the heart ........................................................................... 307
Role of stem cells in cardiac regeneration following injury ................................................... 307
Cardiomyocytes derived from adult skin cells..................................................................... 307
Cardiomyocytes derived from ESCs .................................................................................. 307

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Studies to identify subsets of progenitor cells suitable for cardiac repair ............................... 308
Technologies for preparation of stem cells for cardiovascular therapy ................................... 309
Pravastatin for expansion of endogenous progenitor and stem cells ................................. 309
Cytokine preconditioning of human fetal liver CD133+ SCs ............................................ 309
Expansion of adult cardiac stem cells for transplantation ................................................ 310
Role of MSCs in growth of CSCs .................................................................................. 310
Role of ESCs in repair of the heart ................................................................................... 310
ESC transplantation for tumor-free repair of the heart ................................................... 311
Transplantation of stem cells for myocardial infarction ........................................................ 312
Autologous bone marrow-derived stem cell therapeutics ................................................ 312
Autologous bone marrow-derived mesenchymal precursor stem cells .............................. 312
Transplantation of cord blood stem cells ...................................................................... 312
Transplantation of hESCs ........................................................................................... 313
Transplantation of HSCs ............................................................................................. 313
Transplantation of autologous angiogenic cell precursors................................................ 314
Transplantation of adipose-derived stem cells ............................................................... 314
Transplantation of bone marrow-derived cells for myocardial infarct ................................ 315
Intracoronary infusion of mobilized peripheral blood stem cells ....................................... 316
Transplantation of endothelial cells .............................................................................. 316
Transplantation of cardiomyocytes differentiated from hESCs ......................................... 317
Stem cell therapy for cardiac regeneration ........................................................................ 317
Regeneration of the chronic myocardial infarcts by HSC therapy ..................................... 317
Human mesenchymal stem cells for cardiac regeneration ............................................... 318
In vivo tracking of MSCs transplanted in the heart ........................................................ 319
MSCs for hibernating myocardium ............................................................................... 319
Simultaneous transplantation of MSCs and skeletal myoblasts ........................................ 320
Transplantation of genetically modified cells ................................................................... 320
Transplantation of genetically modified MSCs .................................................................... 320
Transplantation of cells secreting vascular endothelial growth factor ..................................... 320
Transplantation of genetically modified bone marrow stem cells........................................... 320
Cell transplantation for congestive heart failure .............................................................. 320
AngioCell gene therapy for congestive heart failure ............................................................ 321
Injection of adult stem cells for CHF ................................................................................. 322
Intracoronary infusion of cardiac stem cells ....................................................................... 322
Myoblasts for treatment of congestive heart failure ............................................................ 323
Stem cell therapy for dilated cardiac myopathy .................................................................. 323
Role of cell therapy in cardiac arrhythmias ...................................................................... 323
Atrioventricular conduction block ..................................................................................... 324
Genetically engineered cells as biological pacemakers .................................................... 324
Ventricular tachycardia ................................................................................................... 325
Prevention of myoblast-induced arrhythmias by genetic engineering .................................... 325
ESCs for correction of congenital heart defects ................................................................ 326
Cardiac progenitors cells for treatment of heart disease.................................................. 326
Autologus stem cells for chronic myocardial ischemia ..................................................... 327
Role of cells in cardiovascular tissue engineering ............................................................ 327
Construction of blood vessels with cells............................................................................. 327
Engineered arteries for bypass grafts........................................................................... 328
Fetal cardiomyocytes seeding in tissue-engineered cardiac grafts ......................................... 328
Targeted delivery of endothelial progenitor cells labeled with nanoparticles ........................... 328
UCB progenitor cells for engineering heart valves............................................................... 328
Cell therapy for peripheral vascular disease .................................................................... 329
ALD-301 ....................................................................................................................... 329
Cell/gene therapy for PVD ............................................................................................... 329
Cell therapy for CLI in diabetics ....................................................................................... 329
Colony stimulating factors for enhancing peripheral blood stem cells .................................... 330
Intramuscular autologous bone marrow cells ..................................................................... 330
Ixmyelocel-T cell therapy for critical limb ischemia ............................................................. 330
Clinical trials of cell therapy in cardiovascular disease .................................................... 331
Mechanism of the benefit of cell therapy for heart disease .............................................. 333
A critical evaluation of cell therapy for heart disease ...................................................... 334
Publications of clinical trials of cell therapy for CVD ............................................................ 334
Current status of cell therapy for cardiovascular disease ..................................................... 335
Future directions for cell therapy of CVD ......................................................................... 335
Prospects of adult stem cell therapy for repair of heart ....................................................... 336
Combination of cells with biomedical scaffolds ................................................................... 336
Regeneration of cardiomyocytes without use of cardiac stem cells ........................................ 336

6. Cell Therapy for Cancer....................................................................... 339

Introduction..................................................................................................................... 339
Cell therapy technologies for cancer ................................................................................ 339
Cell-based delivery of anticancer therapy ........................................................................ 340

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Cellular immunotherapy for cancer .................................................................................. 340

Treatments for cancer by ex vivo mobilization of immune cells ............................................ 341
Granulocytes as anticancer agents ................................................................................... 342
Neutrophil granulocytes in antibody-based immunotherapy of cancer ................................... 342
Cancer vaccines ............................................................................................................... 342
Autologous tumor cell vaccines ........................................................................................ 342
BIOVAXID ................................................................................................................ 343
OncoVAX .................................................................................................................. 343
Tumor cells treated with dinitrophenyl ......................................................................... 343
Vaccines that simultaneously target different cancer antigens .............................................. 343
Gene modified cancer cells vaccines ................................................................................. 344
GVAX cancer vaccines ................................................................................................ 344
K562/GM-CSF ........................................................................................................... 344
Active immunotherapy based on antigen specific to the tumor ............................................. 345
The use of dendritic cells for cancer vaccination ................................................................. 345
Autologous dendritic cells loaded ex vivo with telomerase mRNA ..................................... 345
Dendritic cell-targeted protein vaccines........................................................................ 346
Dendritic/tumor cell fusion ......................................................................................... 346
Genetically modified dendritic cells .............................................................................. 347
In vivo manipulation of dendritic cells .......................................................................... 347
Preclinical and clinical studies with DC vaccines ............................................................ 347
Vaccines based on dendritic cell-derived exosomes ....................................................... 348
Limitations of DC vaccines for cancer ........................................................................... 348
Future developments to enhance clinical efficacy of DC vaccines ..................................... 348
Lymphocyte-based cancer therapies ................................................................................. 350
Adoptive cell therapy ................................................................................................. 350
Chimeric antigen receptor T cells ................................................................................ 351
Combination of antiangiogenic agents with ACT ............................................................ 352
Expansion of antigen-specific cytotoxic T lymphocytes ................................................... 352
Rescue of CD8+ T cells for use in tumor immunotherapy ............................................... 353
Tumor infiltrating lymphocytes.................................................................................... 353
Genetic engineering of tumor cells to activate T helper cells ........................................... 353
Hybrid cell vaccination .................................................................................................... 354
Chemoimmunotherapy ................................................................................................... 354
Stem cell-based anticancer therapies .............................................................................. 355
Stem cell transplantation in cancer ................................................................................... 355
Peripheral blood stem cell transplantation .................................................................... 355
Stem cell transplantation for hematological malignancies ............................................... 357
Long-term results of HSC transplantation ..................................................................... 358
Prediction of T-cell reconstitution after HSC transplantation. ........................................... 358
HSC transplantation followed by GM-CSF-secreting cell vaccines ..................................... 359
HSC transplantation for renal cell cancer ...................................................................... 359
Complications of stem cell transplants in cancer................................................................. 359
Graft-versus-host disease (GVHD)............................................................................... 359
Delayed immune reconstitution leading to viral infections and relapse ............................. 360
Tumor cell contamination ........................................................................................... 360
Neurological complications ......................................................................................... 361
Hepatic veno-occlusive disease ................................................................................... 361
Current status of the safety of allogeneic HSC transplantation ........................................ 361
Complications of PBSC transplantation in children ......................................................... 362
Role of MSCs in cancer ................................................................................................... 362
MSC-mediated delivery of anticancer therapeutics ......................................................... 362
Nonmyeloablative allogeneic hematopoietic stem cell transplantation ................................... 363
Umbilical cord blood transplant for leukemia...................................................................... 364
hESC-derived NK cells for treatment of cancer ................................................................... 364
ESC vaccine for prevention of lung cancer ......................................................................... 364
Genetic modification of stem cells for cancer therapy.......................................................... 365
Genetic modification of hematopoietic stem cells ........................................................... 365
Use of hematopoietic stem cells to deliver suicide genes to tumors.................................. 365
Delivery of anticancer agents by genetically engineered MSCs ........................................ 365
Mesenchymal progenitor cells for delivery of oncolytic adenoviruses ................................ 366
Genetically modified NSCs for treatment of neuroblastoma............................................. 366
Innovations in cell-based therapy of cancer .................................................................... 367
Use of immortalized cells ................................................................................................ 367
Cancer therapy based on natural killer cells ....................................................................... 367
Cytokine-induced killer cells ............................................................................................ 368
Mesothelin as a target for cancer immunotherapy .............................................................. 368
Nanomagnets for targeted cell-based cancer gene therapy .................................................. 368
Implantation of genetically modified encapsulated cells for anticancer therapy ....................... 369
Antiangiogenesis therapy by implantation of microencapsulated cells............................... 369
Recombinant tumor cells secreting fusion protein .......................................................... 369

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NovaCaps, for pancreatic cancer ............................................................................... 369

A device for filtering cancer and stem cells in the blood ...................................................... 369
Cancer stem cells ........................................................................................................... 370
Role of integrative nuclear signaling in stem cell development ........................................ 370
Cancer stem cell markers ........................................................................................... 371
Breast cancer stem cells ............................................................................................ 371
Role of intestinal stem cells in intestinal polyposis ......................................................... 372
Role of endothelial progenitor cells in tumor angiogenesis .............................................. 372
Role of cancer stem cells in metastases ....................................................................... 372
Therapeutic implications of cancer stem cells ................................................................ 372
Targeting cancer stem cells in leukemia ....................................................................... 373
Targeting stem cells in ovarian cancer ......................................................................... 374
Targeting cancer stem cells to screen anticancer drugs .................................................. 374
Cell-based therapies for malignant brain tumors ................................................................ 374
Bone morphogenetic protein for inhibition of glioblastoma multiforme.............................. 375
Dendritic cell therapy for brain tumors ......................................................................... 375
Encapsulated cells for brain tumors ............................................................................. 376
Immunotherapy of GBM targeting cancer stem cells ...................................................... 376
Mesenchymal stem cells for the treatment of gliomas .................................................... 377
Neural stem cells for drug/gene delivery to brain tumors ............................................... 377
Role of cancer stem cells in resistance to radiotherapy................................................... 378
Targeting stem cells in brain tumors ............................................................................ 379
Companies involved in cell-based cancer therapy ............................................................ 379
American Association for Cancer Research and ESCs ....................................................... 380
Future of cell-based immunotherapy for cancer ............................................................... 381

7. Cell Therapy for Neurological Disorders .............................................. 383

Introduction..................................................................................................................... 383
Regeneration of the nervous system by endogenous stem cells ...................................... 383
Molecular mechanism of neurogenesis .............................................................................. 383
Generation of neurons from astroglia................................................................................ 384
In vivo cell replacement therapy by locally induced neural progenitor cells ............................ 384
Types of cells used for treatment of neurological disorders ............................................. 385
Activated T lymphocytes ................................................................................................. 385
Differentiation of placenta-derived multipotent cells into neurons ......................................... 385
Mesenchymal stem cells induced to secrete neurotrophic factors .......................................... 386
Neural stem cells ........................................................................................................... 386
Development of human CNS stem cells ........................................................................ 386
Distinction between NSCs and intermediate neural progenitors ....................................... 387
Embryonic stem cell-derived neurogenesis ................................................................... 387
Induction of NSCs fro hESCs ....................................................................................... 388
Mechanism of migration of neural stem cells to sites of CNS injury .................................. 389
Monitoring of implanted NSCs labeled with nanoparticles ............................................... 389
Neural progenitor cells ............................................................................................... 390
Neural stem cells as therapeutic delivery vehicles ......................................................... 391
Neural stem cells in the subventricular zone of the brain ................................................ 392
Oligodendrocyte progenitor cells ................................................................................. 392
Promotion of neural stem cells expansion by betacellulin ................................................ 392
Proteomics of neural stem cells ................................................................................... 392
Regulation of neural stem cells in the brain .................................................................. 393
Role of CSF proteins in regulation of neural progenitor cells............................................ 394
Sequencing the transcriptomes of neural stem cells ....................................................... 394
Study of neural differentiation of hESCs by NeuroStem Chip ........................................... 394
Transformation of neural stem cells into other cell types ................................................ 395
Use of epidermal neural crest stem cells for neurological disorders .................................. 395
Olfactory epithelium stem cells for transplantation in the CNS ............................................. 395
Development of CNS cells from non-CNS stem cells ........................................................... 395
Derivation of CNS cells from peripheral nervous system ................................................. 396
Hair-follicle stem cells for neural repair ........................................................................ 397
Stem cells from human umbilical cord blood for CNS disorders ....................................... 397
Immortalized cells for CNS disorders ................................................................................ 397
Fetal tissue transplants ................................................................................................... 398
Choroid plexus cells........................................................................................................ 399
Laboratory mice with human brain cells ............................................................................ 399
Expansion of adult human neural progenitors .................................................................... 399
Neurospheres ................................................................................................................ 400
Dental pulp cells for neuroprotection ................................................................................ 400
Ideal cells for transplantation into the nervous system ....................................................... 400
Cell therapy techniques for neurological applications ...................................................... 401
Carbon nanotubes to aid stem cell therapy of neurological disorders .................................... 401
Cells used for gene therapy of neurological disorders.......................................................... 401

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Fibroblasts ............................................................................................................... 401

Stem cells ................................................................................................................ 402
Neuronal cells ........................................................................................................... 402
Immortalized neural progenitor cells............................................................................ 402
Astrocytes ................................................................................................................ 403
Cerebral endothelial cells ........................................................................................... 403
Human retinal pigmented epithelial cells ...................................................................... 404
Enhancement of growth of stem cells in the brain by drugs ................................................. 404
C3-induced differentiation and migration of NPC for repair of the brain ............................ 405
Stem cell therapies of neurological disorders combined with HBO ......................................... 405
hESCs for CNS repair ...................................................................................................... 405
Methods of delivery of cells to the CNS ............................................................................. 406
Engineered stem cells for drug delivery to the brain ...................................................... 406
Neuronal differentiation of stem cells ................................................................................ 407
Stem cells preparations for CNS disorders ......................................................................... 407
Tracking of stem cells in the CNS by nanoparticles and MRI ................................................ 408
Use of neural stem cells to construct the blood brain barrier ................................................ 409
Encapsulated cells ..................................................................................................... 409
CNS neotissue implant ............................................................................................... 409
Intrathecal delivery of stem cells................................................................................. 410
CNS delivery of cells by catheters ............................................................................... 411
Intravascular administration ....................................................................................... 411
Neurological disorders amenable to cell therapy ............................................................. 411
Neuroprotection by cell therapy ....................................................................................... 412
Cells secreting neuroprotective substances ................................................................... 412
Stem cells for neuroprotection .................................................................................... 412
Neuroprotection by intravenous administration of HSCs ................................................. 412
Human UCB-derived stem cells for the aging brain ........................................................ 413
hESC transplantation to prevent cognitive impairment from radiation .............................. 413
Neurodegenerative disorders ........................................................................................... 413
Role of stem cells in neurodegenerative disorders ......................................................... 413
MSCs for therapy of neurodegenerative disorders.......................................................... 414
Role of NSCs in disorders associated with aging brain .................................................... 415
NSCs for improving memory ....................................................................................... 416
Parkinson's disease ........................................................................................................ 416
Origin and fate of dopamine neurons ........................................................................... 417
Human dopaminergic neurons for PD ........................................................................... 418
Graft survival-enhancing drugs ................................................................................... 418
Xenografting porcine fetal neurons .............................................................................. 419
Encapsulated cells for PD ........................................................................................... 419
Stem cells for PD ...................................................................................................... 419
Stem cells for production of glial derived neurotrophic factor .......................................... 422
Potential of regeneration of endogenous stem cells in PD ............................................... 422
Human retinal pigment epithelium cells for PD .............................................................. 422
Coaxing hESCs to produce dopamine ........................................................................... 423
Tumorigenic potential of transplantated dopaminergic hESCs.......................................... 423
Transplantation of embryonic medial ganglionic eminence cells ....................................... 423
Delivery of cells for PD ............................................................................................... 424
MSCs for multiple system atrophy ............................................................................... 424
Cell therapy for Huntington's disease ................................................................................ 425
Fetal striatal cell transplantation ................................................................................. 425
Transplantation of encapsulated porcine choroids plexus cells ......................................... 425
Cell therapy for Alzheimer's disease ................................................................................. 426
Neural stem cell implantation for Alzheimer's disease .................................................... 426
Implantation of genetically engineered cells producing NGF ............................................ 426
Cell therapy for amyotrophic lateral sclerosis ..................................................................... 427
Stem cell techniques for study of ALS .......................................................................... 427
Use of stem cells for ALS............................................................................................ 428
Transplantation of glial restricted precursors in ALS ....................................................... 429
Stem cell-based drug discovery for ALS ....................................................................... 430
Cell therapy for demyelinating disorders ........................................................................... 430
Autologous bone marrow stem cell therapy for multiple sclerosis .................................... 431
ESCs for remyelination .............................................................................................. 431
Fusokine method of personalized cell therapy of MS ...................................................... 431
Hematopoietic stem cell transplantation for MS ............................................................. 432
Mechanism of repair of demyelination after NSC transplantation ..................................... 432
MSCs for multiple sclerosis ......................................................................................... 433
Neural progenitor cells for neuroprotection in MS .......................................................... 433
T cell-based personalized vaccine for MS ...................................................................... 434
Stem cells for chronic inflammatory demyelinating polyneuropathy ................................. 434
X-linked adrenoleukodystrophy ................................................................................... 434

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Cell therapy of stroke ..................................................................................................... 434

Adult stem cell therapy in stroke ................................................................................. 435
Implantation of genetically programmed ESCs .............................................................. 436
Intravenous infusion of MSCs ..................................................................................... 436
Intravenous infusion of human UCB stem cells .............................................................. 437
Intracerebral administration of human adipose tissue stromal cells.................................. 438
Neural stem cell therapy for stroke .............................................................................. 438
Transplantation of encapsulated porcine choroids plexus ................................................ 439
Transplantation of fetal porcine cells ............................................................................ 439
Role of cell therapy in management of stroke according to stage .................................... 440
Clinical trials of cell therapy for stroke ......................................................................... 440
Future of cell therapy for stroke .................................................................................. 441
Cell therapy of traumatic brain injury ............................................................................... 442
Cell/gene therapy for TBI ........................................................................................... 443
Clinical trials of autologous HSC therapy for TBI............................................................ 443
Limitations of stem cell therapy for acute TBI ............................................................... 444
Improving the microenvironments of transplanted cells in TBI ........................................ 444
Cell therapy for spinal cord injury .................................................................................... 444
Autoimmune T cells against CNS myelin-associated peptide ........................................... 445
Fetal neural grafts for SCI .......................................................................................... 445
Olfactory-ensheathing cells for SCI.............................................................................. 445
Oligodendrocyte precursor cells for treatment of SCI ..................................................... 446
Schwann cell transplants for SCI ................................................................................. 446
Transplantation of glial cells for SCI ............................................................................ 446
Stem cells for SCI .......................................................................................................... 446
Bone marrow stem cells for SCI .................................................................................. 447
Embryonic stem cells for SCI ...................................................................................... 447
Transplantation of induced pluripotent stem cells in SCI................................................. 448
Transplantation of MSCs for SCI.................................................................................. 448
Transplantation of NSCs for SCI .................................................................................. 448
Transplantation of human dental pulp stem cells ........................................................... 449
Transdifferentiation of BM stem cells into cholinergic neurons for SCI .............................. 449
Evaluation of experimental studies of stem cell transplantation in SCI ............................. 450
Spinal stem cells for treatment of ischemic injury of spinal cord ........................................... 450
Combined approaches for regeneration in SCI ................................................................... 451
Combined cell/gene therapy for SCI ............................................................................ 451
Delivery of cells in SCI .................................................................................................... 451
Intrathecal injection of cells labeled with magnetic nanoparticles .................................... 452
Intravenous injection of stem cells for spinal cord repair ................................................ 452
Clinical applications of stem cells for SCI........................................................................... 452
Autologous bone marrow cell transplantation for SCI ..................................................... 452
Cell therapy of syringomyelia...................................................................................... 453
Cell therapy for neurogenetic disorders ............................................................................. 453
Hurler's syndrome treated with stem cells .................................................................... 453
Krabbes disease treated with UCB stem cells ............................................................... 454
Krabbe's disease treated with combination of cell and gene therapy ................................ 454
Mitochondrial encephalomyopathies treated with stem cells ............................................ 455
Sanfilippo syndrome type B treated with UCB stem cells ................................................ 455
Cell therapy for lysosomal storage disorders ...................................................................... 455
Cell therapy for Batten disease ................................................................................... 455
Cell/gene therapy for Farbers disease ......................................................................... 456
Genetically modified HSCs for metachromatic leukodystrophy ......................................... 456
Neural stem cells for lysosomal storage disorders.......................................................... 457
Cell therapy of epilepsy .................................................................................................. 457
Cell therapy of posttraumatic epilepsy ......................................................................... 457
Cell therapy for temporal lobe epilepsy ........................................................................ 458
Cell therapy for pharmacoresistant epilepsies ............................................................... 458
Cell therapy for developmental neurological disorders......................................................... 459
Cell therapy for cerebral palsy ......................................................................................... 459
Cell therapy for muscle disorders ..................................................................................... 459
Duchenne muscular dystrophy .................................................................................... 460
Combination of cell and pharmacotherapy for DMD........................................................ 460
Myoblast transplant for DMD ...................................................................................... 460
Myoblast-based gene transfer ..................................................................................... 461
Myoblasts lacking the MyoD gene ................................................................................ 461
Myoblast injection for treatment of other muscular dystrophies ....................................... 462
Role of satellite cells in the treatment of DMD ............................................................... 462
Stem cells for DMD.................................................................................................... 463
Cell therapy for autism ................................................................................................... 464
Management of chronic intractable pain by cell therapy ...................................................... 464
Implantation of chromaffin cells .................................................................................. 465

- 16 -

Role of stem cells in management of pain .................................................................... 465

Implantation of astrocytes secreting enkephalin ............................................................ 466
Cells for delivery of antinociceptive molecules ............................................................... 466
Implantation of genetically engineered cells ................................................................. 466
Cell therapy for low back pain ..................................................................................... 466
Cell therapy for neuroendocrine disorders ......................................................................... 467
Pituitary stem cells .................................................................................................... 467
Cell therapy for retinal degenerative disorders ................................................................... 467
Human retinal stem cells ............................................................................................ 468
Delivery of CNTF by encapsulated cell intraocular implants ............................................. 469
Stem cell transplantation in the retina ......................................................................... 469
ESCs for retinal degenerative disorders ........................................................................ 470
hESC-derived RPE cells for macular dystrophy .............................................................. 470
Neuroprotective effect of neural progenitor cell transplantation ....................................... 470
Genetically engineered retinal pigmented epithelial cell lines .......................................... 471
Combining cell and gene therapies for retinal disorders .................................................. 471
Stem cell therapy for hearing loss .................................................................................... 471
Cell thery for peripheral nerve lesions ............................................................................... 472
Cell transplants for peripheral nerve injuries ................................................................. 472
Treatment of diabetic neuropathy with endothelial progenitor cells .................................. 472
Complications of cell therapy of CNS disorders ................................................................ 472
Tumor formation after CNS transplantation of stem cells ..................................................... 473
Uncontrolled differentiation of implanted cells ............................................................... 473
Donor stem cell-derived brain tumor ........................................................................... 473
Clinical trials of cell therapy in neurological disorders ..................................................... 473
Future prospects for cell therapy of CNS disorders .......................................................... 475

8. Ethical and Political Aspects of Cell therapy........................................ 477

Introduction..................................................................................................................... 477
Political and ethical aspects of hESC research in the US .................................................. 477
Ethical issues concerning fetal tissues ............................................................................... 477
Morality and hESC research ........................................................................................ 477
Opponents of hESC research in the US ............................................................................. 478
Use of hESCs in NIH-supported research ...................................................................... 479
Politics of hESC research in the US .............................................................................. 480
Public opinion in the US about hESC research ............................................................... 482
Human stem cell cloning in the US .............................................................................. 483
Stem cell guidelines of various US institutions .............................................................. 484
Ethics of transplanting human NSCs into the brains of nonhuman primates ........................... 484
Stem cell lines available worldwide ................................................................................. 485
Stem cell policies around the world ................................................................................. 486
Countries with no defined policies on hESC research........................................................... 486
Australia ....................................................................................................................... 487
Canada ......................................................................................................................... 487
China ........................................................................................................................... 488
Denmark....................................................................................................................... 489
France .......................................................................................................................... 489
Germany....................................................................................................................... 489
India ............................................................................................................................ 491
Ireland ......................................................................................................................... 492
Israel ........................................................................................................................... 492
Italy ............................................................................................................................. 492
Japan ........................................................................................................................... 493
The Netherlands ............................................................................................................ 493
Saudi Arabia.................................................................................................................. 494
Singapore ..................................................................................................................... 494
South Africa .................................................................................................................. 494
South Korea .................................................................................................................. 495
Spain ........................................................................................................................... 495
Sweden ........................................................................................................................ 495
Switzerland ................................................................................................................... 496
United Kingdom ............................................................................................................. 496
ESC bank ................................................................................................................. 497
European Union ............................................................................................................. 498
EU guidelines for stem cell research ............................................................................ 498
EMBOs recommendations for stem cell research ........................................................... 499
Public opinion in Europe about hESC research.................................................................... 500
United Nations, cloning and nuclear transfer ...................................................................... 501
The Embryo Project for information on ESC research .......................................................... 502
Concluding remarks about ethics of ESC research .............................................................. 502
Ethical issues concerning umbilical cord blood ................................................................ 502

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Cell therapy tourism ........................................................................................................ 503

9. Safety and Regulatory Aspects of Cell Therapy ................................... 505

Introduction..................................................................................................................... 505
Safety issues of cell therapy ............................................................................................ 505
Immune-mediated reactions to transpanted stem cells ....................................................... 505
Human virus infections associated with stem cell transplantation ......................................... 506
Herpes simplex virus type 1 ....................................................................................... 506
Cytomegalovirus ....................................................................................................... 506
Opportunistic infections among hematopoietic stem cell transplant recipients ........................ 506
Cord colitis syndrome ..................................................................................................... 506
Carcinogenic potential of stem cells and its prevention ....................................................... 507
FDA safety regulations for cell and tissue products ......................................................... 507
FDA Guidance on license applications for umbilical cord blood products ................................. 508
Regulation of cord blood banks in the US ......................................................................... 508
Regulatory issues for biotechnology-derived drugs ......................................................... 508
Regulation of cell selection devices for PBSCs at point of care......................................... 509
FDA rules for human cells and tissues ............................................................................. 510
FDA regulation of fetal cellular or tissue products ............................................................... 510
FDA and clinical trials using hESCs ................................................................................... 511
Cell and gene therapy INDs placed on hold by the FDA ....................................................... 511
Regulatory issues for genetically engineered cell transplants ............................................... 512
FDA guidelines for human tissue transplantation ................................................................ 512
Xenotransplantation ........................................................................................................ 512
Clinical Protocol Review and Oversight .............................................................................. 513
Informed consent and patient education ........................................................................... 513
Xenotransplantation product sources ................................................................................ 513
FDA guidelines for xenografts .......................................................................................... 514
Regulations relevant to cell therapy in the European Union ............................................. 515
Regulations about use of stem cells in the EU .................................................................... 516
EMEA regulation of cell/gene therapy ............................................................................... 516
Guidelines for cell therapy in the UK ................................................................................. 517
NIH and stem cells ........................................................................................................... 518
hESC lines approved under the new NIH guidelines ............................................................ 518
Clinical trials in cell therapy ............................................................................................. 519
Stem cell patents ............................................................................................................. 519
Stem cell patents in the United States .............................................................................. 519
Current status of Thomson patents at WARF ................................................................. 520
Stem cell patents in the European Union ........................................................................... 520

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1-1: Landmarks in the history of cell therapy ........................................................................ 27

1-2: Examples of cells involved in various diseases................................................................ 32
2-1: Types of human cells used in cell therapy ...................................................................... 35
2-2: A selection of companies providing cell culture media ..................................................... 39
2-3: A sampling of companies supplying cell sorters .............................................................. 44
2-4: Companies involved in cell-based drug discovery ........................................................... 51
2-5: Methods of delivery of cells for therapeutic purposes ...................................................... 53
2-6: Therapeutic applications of encapsulated cells ................................................................ 57
2-7: Companies working on encapsulated cell technology ....................................................... 60
2-8: Molecular imaging methods for tracking cells in vivo ....................................................... 65
3-1: Companies involved in cord blood banking as a source of stem cells ............................... 101
3-2: Sources of adult human stem cells ............................................................................. 112
3-3: Comparison of human stem cells according to derivation ............................................... 121
3-4: Enhancing engraftment, mobilization and expansion of stem cells .................................. 142
3-5: Applications of stem cells .......................................................................................... 158
3-6: Advantages and limitations of methods for optimizing MSCs .......................................... 168
3-7: Growth factors with positive effects on stem cells and applications ................................. 176
3-8: Examples of drugs that induce granulocytopenia at stem cell level ................................. 186
3-9: Academic institutes involved in stem cell research ........................................................ 193
3-10: Companies involved in stem cell technologies ............................................................ 195
4-1: Therapeutic applications of regulatory T cells (T-regs) .................................................. 212
4-2: Various tissue/cell therapy approaches to the treatment of type 1 diabetes ..................... 218
4-3: Companies involved in cell therapy for insulin-dependent diabetes ................................. 234
4-4: Major pulmonary disorders potentially treatable by stem cell manipulation ...................... 250
4-5: Cell-based repair of knee cartilage damage ................................................................. 267
5-1: Classification of various types of cell therapy for cardiovascular disorders ....................... 295
5-2: Clinical trials of cell therapy in cardiovascular disease ................................................... 331
6-1: Cell therapy technologies used for cancer .................................................................... 339

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Companies involved in developing cell-based therapies for cancer .................................. 379

Experimental use of immortalized cells for CNS disorders .............................................. 398
Combination of stem cells and HBO in models of neurological disorders .......................... 405
Methods for delivering cell therapies in CNS disorders ................................................... 406
Neurological disorders amenable to cell therapy ........................................................... 411
Types of cell used for investigative treatment of Parkinson's disease............................... 416
Status of cell therapies for Parkinson's disease............................................................. 417
Role of cell therapy in management of stroke according to stage .................................... 440
Clinical trials of cell therapy for stroke: completed, ongoing and pending ........................ 440
Clinical trials with cell-based therapies in neurological disorders (excluding stroke) .......... 474
Listed numbers of stem cell lines around the world as of end of 2008 ............................. 485
Stem cell policies around the world............................................................................. 486
European public attitudes about research involving human stem cells ............................. 501
Possible adverse reactions and safety issues of cell therapy ........................................... 505

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1-1:
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5-1:
6-1:
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Interrelationships of cell therapy to other technologies .................................................. 29

Interrelationships of gene, cell and protein therapies ..................................................... 31
A simplified biological scheme of embryonic stem Cells .................................................. 72
Steps of iPS cell production ...................................................................................... 105
Flow chart of development of stem cells with potential bottlenecks ................................ 200
hESC-derived cardiomyocytes from laboratory to bedside ............................................ 308
A scheme of generation and administration of tumor antigen-pulsed dendritic cells ......... 349
Stem cell transplantation techniques ......................................................................... 356
Stem cells that can give rise to neurons ..................................................................... 391
Approaches to stem cell therapy in stroke .................................................................. 436

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