Am J Physiol Gastrointest Liver Physiol 300: G684G696, 2011.

Review First published January 20, 2011; doi:10.1152/ajpgi.00474.2010.

Intestinal myofibroblasts: targets for stem cell therapy

R. C. Mifflin,1 I. V. Pinchuk,1 J. I. Saada,1 and D. W. Powell1,2
Departments of 1Internal Medicine and 2Neuroscience and Cell Biology, University of Texas Medical Branch,
Galveston, Texas
Submitted 19 October 2010; accepted in final form 17 January 2011

Mifflin RC, Pinchuk IV, Saada JI, Powell DW. Intestinal myofibroblasts: targets
for stem cell therapy. Am J Physiol Gastrointest Liver Physiol 300: G684G696, 2011.
First published January 20, 2011; doi:10.1152/ajpgi.00474.2010.The subepithelial in-
testinal myofibroblast is an important cell orchestrating many diverse functions in
the intestine and is involved in growth and repair, tumorigenesis, inflammation, and
fibrosis. The myofibroblast is but one of several -smooth muscle actin-positive

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(-SMA) mesenchymal cells present within the intestinal lamina propria, includ-
ing vascular pericytes, bone marrow-derived stem cells (mesenchymal stem cells or
hematopoietic stem cells), muscularis mucosae, and the lymphatic pericytes (colon)
and organized smooth muscle (small intestine) associated with the lymphatic
lacteals. These other mesenchymal cells perform many of the functions previously
attributed to subepithelial myofibroblasts. This review discusses the definition of a
myofibroblast and reconsiders whether the -SMA subepithelial cells in the
intestine are myofibroblasts or other types of mesenchymal cells, i.e., pericytes.
Current information about specific, or not so specific, molecular markers of lamina
propria mesenchymal cells is reviewed, as well as the origins of intestinal myofi-
broblasts and pericytes in the intestinal lamina propria and their replenishment after
injury. Current concepts and research on stem cell therapy for intestinal inflam-
mation are summarized. Information about the stem cell origin of intestinal stromal
cells may inform future stem cell therapies to treat human inflammatory bowel
disease (IBD).
pericytes; bone marrow stem cells; hematopoietic stem cells; mesenchymal stem
cells; mesenchymal cells; stromal cells; muscularis mucosae; lymphatic lacteal;
-smooth muscle actin; thymic antigens; injury and repair; inflammatory bowel
disease; Crohns disease; bone marrow transplantation; Thy-1; NG2; fibroblast-
specific protein 1; patched; periostin; fibroblast activation protein; epimorphin;
bone marrow transplantation

IN THE PAST DECADE, the subepithelial intestinal myofibroblast
has come to be regarded as an important cell orchestrating
many diverse functions in the intestine in health and disease,
including growth and repair, tumorigenesis and cancer progres-
sion, inflammation or peripheral immune tolerance, and fibro-
sis. These concepts have been summarized in several thought-
ful and informative reviews (5, 21, 33, 46, 9799). Our own
research indicates that the intestinal myofibroblast is but one of
several mesenchymal cells in the intestinal lamina propria,
and many of the functions attributed to subepithelial myo-
fibroblasts might be due to other -smooth muscle actin-
positive (-SMA) mesenchymal cells such as pericytes,
bone marrow-derived stem cells [mesenchymal stem cells
(MSCs) or hematopoietic stem cells (HSCs)], muscularis
mucosae, and the lymphatic pericytes (colon) and organized
smooth muscle (small intestine) associated with the lym-
phatic lacteals (93, 99).
There are three goals for this review. The first is to review
the definition of a myofibroblast and reconsider whether the
Address for reprint requests and other correspondence: D. W. Powell, Depts.
of Internal Medicine and Neuroscience and Cell Biology, 4.106 McCullough
0764, 301 University Blvd., Galveston, TX 77555-0764 (e-mail: dpowell
@utmb.edu).
G684 0193-1857/11 Copyright 2011 the American Physiological Society
-SMA subepithelial cells in the intestine are myofibroblasts
or other types of mesenchymal cells, i.e., pericytes.
A second goal is to compile current information about
specific, or not so specific, molecular markers of lamina pro-
pria mesenchymal cells. Because advances in our understand-
ing of the role of these cells in health and disease depend on
our ability to identify them in human and animal tissues and to
ablate them in disease models, we review here the marker
molecules that might be useful in this regard.
The third goal is to review our current understanding of
the origins of intestinal myofibroblasts and pericytes in the
intestinal lamina propria and their replenishment after in-
jury. Tissue stromal cells have been postulated to play a
fundamental role in tissue regeneration and wound repair
(112), and stem cell transplantation has been shown to repair
the inflamed intestine (35). Information about the stem cell
origin of intestinal stromal cells may inform attempts to
develop stem cell therapies to treat human inflammatory
bowel disease (IBD).
Myofibroblasts
Eyden (30) has challenged the idea that the subepithelial
cells in the intestine are true myofibroblasts, and, following a
review of our own histological material and the literature, we
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MYOFIBROBLASTS AND STEM CELL THERAPY
believe he is correct in questioning this concept. Eyden defines
myofibroblasts in detail and with strict criteria: spindle-shaped
or stellate cells that are -SMA positive, vimentin positive,
smooth muscle myosin negative but non-smooth muscle myo-
sin positive, fibronectin positive, and very weakly positive or
negative for desmin. On electron microscopy (EM), the cells
possess abundant cytoplasm and pericellular matrix and con-
tain a prominent rough endoplasmic reticulum and a Golgi
apparatus that produces collagen granules. The cells are at-
tached to adjacent myofibroblasts by gap junctions and spe-
cialized adherens junctions containing OB-cadherin (47) and,
most importantly, are attached to the surrounding matrix
through a fibronexus and not basal lamina. The fibronexus is an
adhesion structure connecting the myofibroblast to the matrix
through extracellular fibronectin filaments that run parallel to
and then cross into the cytoplasm of the myofibroblast, forming
a plaquelike structure where fibrils enter the cell membrane.
These adhesions to matrix and to each other make them a
powerful contractile network, which might have a teleolog-
ical purpose of reducing the size of a wound. According to
this strict definition, true myofibroblasts would be rele-
gated to wound-healing granulation tissue and the stroma of
neoplasms (30).
Pericytes
Pericytes, also called mural cells and vascular smooth mus-
cle cells, were described over 100 years ago. EM and immu-
nohistochemical studies have shown them to be cells with a
prominent nucleus and long processes aligned longitudinally
along the axis of the vessel with smaller, thin circumferential
processes that partially engirdle the vascular wall. In histolog-
ical sections these processes might not be seen, giving the cells
a fusiform morphology like myofibroblasts (10, 67). The mural
cells are embedded within and contribute to the formation of
the basement membrane of microvessels. Gap junctions di-
rectly connect pericytes with each other and with the underly-
ing endothelial cells, and adhesion plaques to matrix augment
this attachment. The cells stain positive for -SMA, desmin,
melanoma chondroitin sulfate proteoglycan (MCSP, also
called NG2, see Melanoma chondroitin sulfate proteoglycan
below), PDGF receptor (PDGFR)-, and regulator of G protein
signaling-5 (RGS5). Pericytes also express monocyte-mac-
rophage markers such as CD11b, and, upon stimulation with
interferon (IFN)-, they express major histocompatibility com-
plex (MHC) class II molecules and B7 family costimulatory
molecules CD80 and CD86, suggesting that they are replen-
ished by circulating fibrocytes (see Hematopoietic stem cells or
fibrocytes below). The ability of cultured fibrocytes to support
angiogenesis/vasculogenesis by cultured endothelial cells en-
hances this idea (42), since angiogenesis is known to require
both endothelial cells and pericytes. Pericytes also express
receptors for and respond to both cholinergic and adrenergic
agents, angiotensin II and endothelin-1 (ET-1) (as do cultured
subepithelial myofibroblasts).
Hepatic stellate cells (Ito cells) and renal mesangial cells are
examples of pericytes, and pericytes are prominent in the
stroma of neoplasms, where they probably play a role in tumor
angiogenesis (10).
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G685
Organization of Intestinal Mesenchymal (Stromal) Cells
-SMA subepithelial cells. The subepithelial fibroblast-like
mesenchymal cells were first defined as fibroblasts or reticu-
lohistiocytic cells (22, 27, 68). Between 1968 and 1971, a
series of papers described this organelle in the colon and
named it the pericryptal fibroblastic sheath: a group of fusi-
form-shaped fibroblasts embedded in collagen and mucopoly-
saccharide ground substance immediately subjacent to the
epithelial basement membrane, encircling the colonic crypts
and ending in the collagen table just under the colonic surface
epithelium. In the basal one-third of the crypts, the fibroblast
sheath was observed to be two or three cells thick, often
overlapping like shingles. In this location, the cells did not
appear to contain a complex cytoplasm with active Golgi. At
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the mouth of the crypts, the layer is one cell thick and more
sparse and fenestrated (stellatelike), and the cells demonstrate
long cytoplasmic processes with a rich endoplasmic reticulum
and Golgi apparatus (55, 56, 90).
Marsh and Trier defined the anatomy of the subepithelial
sheath in the mouse small intestine, noting that the fibroblastic
sheath surrounding the small intestinal crypts has an appear-
ance similar if not identical to the surrounding colonic crypts.
They reported (76, 77) the lack of a well-formed sheath in the
villi, where little pericellular collagen was found and the
fibroblasts seemed associated with endothelial and immune
cells. Using 3H-methyl-thymidine as a tracer of mitotic activ-
ity, studies in the rabbit colon (90) and mouse small intestine
(77) suggested that the fibroblasts of the sheath divided first in
the crypt region and migrated up the villus like the epithelium.
These observations suggested that lamina propria fibroblasts,
like the epithelium, were replaced by dividing stem cells.
Subsequent similar studies in mouse intestine and colon failed
to confirm this idea, as thymidine-labeled cells were seen
randomly in the subepithelial sheath (83). These labeling
studies do indicate a capability of the subepithelial fibroblast-
like cells to divide and contribute to their stability as an
organelle in the intestine.
In 1987, Joyce et al. (51) performed a detailed EM study of
the lamina propria of rat small intestine and colon, confirming
the anatomy of the crypt area described by previous authors but
noting the stellate appearance of the villus subepithelial cells
whose processes connected to the matrix with adherence
plaques and through adherens junctions to each other, to crypt
subepithelial cells, and often to a third dimension of cells in the
lamina propria. These cells were weakly SMA, tropomyosin,
and both smooth muscle and non-smooth muscle myosin pos-
itive (although less so than the smooth muscle of the muscu-
laris mucosae and lymphatic lacteal), indicating their contrac-
tile capability. Furuya and Furuya (33) also performed ele-
gant histological studies and demonstrated the contractile
nature of this network, confirming with elegant studies its
mechanosensitive properties, the connections of the network
through dye-permeable junctions, and its contractile re-
sponse to ET-1 and purinergic transmitters. The report of
Joyce et al. and subsequent EM studies confirmed the
anatomy of this subepithelial organelle (124). -SMA im-
munohistochemistry led to labeling these subepithelial cells
in the colon as myofibroblasts (2, 98). In the upper aspect of
the crypts and subsurface epithelial region of the colon, the
subepithelial cells lose -SMA staining and become more
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G686 MYOFIBROBLASTS AND STEM CELL THERAPY
Fig. 1. -Smooth muscle actin (-SMA)-positive cells of the colonic mucosa.
Cross section (A) and longitudinal sections (B and C) of the colonic mucosa
show positive (brown) -SMA immunostaining of pericryptal (subepithelial)
myofibroblasts (white arrows) and the muscularis mucosae (stars). Vascular
and/or lymphatic pericytes (white arrowheads) are also visible. Original
magnification 200. C: pericytes surrounding lymphatics (black arrows) are
seen entering the muscularis mucosae in this section. Note that the lamina
propria blood vessels between the crypts and muscularis mucosae are also
highlighted (original magnification 400). [A and B taken from Powell et al.
(96), with permission. C adapted from Adegboyega et al. (3), with permission].
stellate in shape, i.e., become fibroblasts or stellate cells.
Deeper in the core of the colonic lamina propria, some of the
-SMA cells are clearly pericytes surrounding microves-
sels and smaller lymphatic vessels as they make their way to
and through the muscularis mucosae (Fig. 1).
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In the small intestinal villus (Fig. 2), -SMA staining of
subepithelial cells is less than those surrounding the crypts, and
may even be lost altogether in the mid- and upper villus. These
cells stain weakly for desmin, suggesting that they might be
pericytes. Scanning EM of the small intestinal villus after freeze
fracture and osmic acid, HCl, NaOH, or enzymatic digestion
reveals a subepithelial basement reticular network with embedded
fibroblasts that surround fenestrae in the matrix (23, 24, 61, 62,
121). It is likely that these fenestrae are actually the digested
remnants of subepithelial microvessels that abut onto the epithe-
lial basement membrane. In scanning EM studies where the villus
subepithelial fibroblasts are intact, it is clear that they overlie
microvessels (23). Thus we conclude that the predominant
-SMA subepithelial cells in the intestinal villus are pericytes,
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although rare myofibroblasts may also be present.
We now believe that in the crypts of the colon and small
intestine the subepithelial -SMA cells are activated fibro-
blasts (myofibroblasts), which are connected by gap or adher-
ens junctions to a second or sometimes third layer of -SMA
fibroblasts to form a sheath that is located in the lamina propria
surrounding the crypt epithelium. These cells appear to be
different from -SMA vascular and lacteal pericytes within
the lamina propria, but three-dimensional confocal or mul-
tiphoton microscopic techniques will be necessary to rule out
the possibility that they might also surround vessels in some
unseen dimension, and thus might actually be pericytes. Al-
though all of Edyens criteria have not been satisfied, these
-SMA- and vimentin-positive, desmin-negative cells are not
fibroblasts, are probably not pericytes, and are not smooth
muscle cells, and so with some license, subepithelial myofi-
broblast seems to be a good name for this first layer of
Fig. 2. -SMA-positive cells of the small intestinal mucosa. A: longitudinal
section of normal human jejunum with positive (brown) -SMA immunostain-
ing of myofibroblasts (white arrows), pericytes (white arrowheads), lymphatic
lacteal-associated smooth muscle (black arrows), and muscularis mucosae
(stars). B: magnified image of box shown in A demonstrating intensely
-SMA-positive smooth muscle (black arrows) associated with the lymphatic
lacteal and mildly -SMA-positive cells (white arrowheads) subjacent to the
epithelium. These subepithelial cells appear to be predominantly pericytes
rather than myofibroblasts. [Courtesy of Patrick Adegboyega, Louisiana State
University Health Sciences Center (Shreveport, LA). Adapted from Powell et
al. (99) with permission.]
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MYOFIBROBLASTS AND STEM CELL THERAPY
pericryptal mesenchymal cells. Furthermore, fibronectin im-
munostaining has not been performed to seek fibronexi asso-
ciated with these cells, which might better justify naming them
myofibroblasts, although adhesion plaques to underlying ma-
trix have been reported (51).
Intestinal fibroblasts. Intestinal fibroblasts are currently rec-
ognized by their morphology and the absence of -SMA
staining. In the colonic mucosa, they may be found in the
lamina propria as a second layer of cells adjacent to the
myofibroblasts surrounding the colonic crypts. The predomi-
nant collection of fibroblasts is in the upper aspects of the
colonic crypts, especially in the region under the surface
epithelium where the fibroblastic sheath is attenuated and
fenestrated. Another area very rich in fibroblasts is the subep-
ithelial region of the middle and upper portions of the small
intestinal villus (33). Fibroblasts in many tissues are identified
by immunostaining for CD90 (Thy-1) (see Thymus stromal
antigens below). However, both Thy-1-positive and Thy-1-
negative fibroblasts are reported in the human myometrium and
orbit of the eye, and each type shows a fundamentally different
response to signaling molecules. For example, only Thy-1-
positive orbit and uterine fibroblasts express -SMA in re-
sponse to transforming growth factor (TGF)- (i.e., become
myofibroblasts), while only Thy-1-negative cells respond to
15-deoxy-PGJ2 with lipid storage (i.e., become stellate cells)
(66). Of interest is the observation that cultured intestinal
myofibroblasts assume a stellate morphology in response to
elevated intracellular cyclic AMP (124).
Figure 3 is a schematic drawing illustrating our current
concepts of the subepithelial cells of the colonic and small
intestinal crypts and small intestinal villus. This drawing dif-
fers from our previous renditions, pointing out our understand-
ing that only the first layer of subepithelial mesenchymal cells
in the colonic crypts are myofibroblasts and that subepithelial
cells in the villus of the small intestine are mostly pericytes.
These -SMA cells, together with -SMA fibroblasts, rep-
resent the stromal cells of the intestinal lamina propria.
Identification of Intestinal Stromal Cells
We believe that the subepithelial mesenchymal cells are
critical to the initiation and perpetuation of intestinal inflam-
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G687
mation and to the therapy of IBD. Either effective therapy may
come through medications that inhibit mesenchymal cell-de-
rived inflammatory mediators or processes or, alternatively,
novel therapies might be agents that might mimic or promote
the salutary mediators and processes (tolerance) for which
these stromal cells may be responsible. In light of the effect of
anti-TNF antibodies in rheumatoid arthritis, which is directed
toward the joint synoviocyte, also an antigen-presenting myo-
fibroblast, it is not unreasonable to propose that the beneficial
effects of such therapy in IBD are in part derived from
anti-inflammatory effects on intestinal stromal cells, which
also possess antigen presenting capability (92, 105). Further-
more, a TNF-overexpressing mouse model develops both
chronic inflammatory polyarthritis and Crohns-like IBD (63)
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that is dependent on TNF receptor (TNFR)I expression in
synovial myofibroblasts and intestinal myofibroblasts (7).
To more definitively study the normal biology of intestinal
mesenchymal cells, and to understand their role in human
disease, we must be able to identify these cells and ablate them
in experimental animal models. Intestinal mesenchymal cells
are usually identified by using a panel of molecular markers,
hoping to distinguish one cell type from another. However, this
approach is rarely definitive given the plasticity of mesenchy-
mal cells and the significant overlap of marker expression
among the different cells. For example, desmin expression is
often used to distinguish myofibroblasts from smooth muscle,
yet pericytes express desmin. Below we discuss some of the
more useful markers employed to distinguish intestinal mes-
enchymal cells. It should become clear that while many of
these markers may be useful in the identification of various
intestinal mesenchymal cells, none is capable of exclusively
defining each population.
-Smooth muscle actin. Myofibroblasts have been identified
by the expression of -SMA, vimentin, or desmin, type I
collagen maturation enzymes such as prolyl-4-hydroxylase
(P4H), and the absence of epithelial cytokeratins (21). -SMA
remains the best single marker of the subepithelial myofibro-
blast network, but it does not differentiate myofibroblasts from
the other mesenchymal elements of the lamina propria; i.e.,
mural cells (pericytes) of the lamina propria vessels (9, 96),
bone marrow-derived (produced) lamina propria stromal stem
Fig. 3. Schematic depicting the spatial rela-
tionships of the epithelium and mesenchymal
elements of the small intestinal villus and
colonic crypts. A: cross section through a
small intestinal villus showing the epithelium
(Epi) and lamina propria. -SMA subepithe-
lial cells are predominantly pericytes (P,
green) surrounding small blood vessels (BV).
There may be occasional myofibroblasts
(MF), although these could all be pericytes in
the villus. There are smooth muscle (SM)
bundles (dark green) associated with the cen-
tral lymphatic lacteal (CL). Fibroblasts (FB,
gray) are shown. B: cross section through the
colonic crypts demonstrating relationships
among the various mesenchymal elements in
the lamina propria: subepithelial myofibro-
blasts (MF, green) and pericytes (P, green)
surrounding blood vessels (BV) and lymphat-
ics (L). Fibroblasts (F, gray) are found deeper
in the lamina propria. EC, endothelial cell.
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G688 MYOFIBROBLASTS AND STEM CELL THERAPY

cells (9, 16), the muscularis mucosae (2), and the smooth
muscle of the small intestinal villus core (75). The smooth
muscle fibers of the small intestinal villus core are likely the
lymphatic lacteals (Fig. 2), a conclusion supported by compar-
ing -SMA-stained histological sections with published scan-
ning electron micrograph of corrosion casts of intestinal lacteals
(86) and immunohistochemical sections stained with antibodies
against the lymphatic endothelium (LYVE1, a hyaluronan recep-
tor) (74).
Intermediate filament stains differentiate the myofibroblasts
from smooth muscle of the muscularis mucosae or the lym-
phatic lacteal, the myofibroblasts staining strongly for vimentin
and the smooth muscle staining strongly for desmin (2, 3, 75).
-SMA is downregulated by IFN- (25, 66), and thus myofi-
expressed by subsets of fibroblasts, myofibroblasts, vascular
pericytes, HSCs and MSCs, activated endothelial cells, and
some murine neurons and murine T cells (which limits use of
this marker in murine studies) (12, 100, 101). Antibodies
against this surface protein identify the human intestinal myo-
fibroblast-fibroblast-pericyte network, where diffuse staining
suggests that the extracellular matrix (ECM) may also capture
soluble Thy-1 (94).
Intestinal fibroblasts, myofibroblasts, pericytes, and lym-
phatic stromal cells can also be identified with monoclonal
antibodies raised against thymic stromal cells such as ER-TR7
(53, 54, 84, 126) and TE-7 (36, 44). Preliminary studies in our
laboratory show reactivity of Thy-1, ER-TR7, and -SMA
against these mesenchymal elements of the lamina propria, and

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broblasts may be overlooked in inflamed tissues. confocal microscopic merged images (Fig. 4) give promise that
Thymus stromal antigens. Thy-1 (CD90) is a 25- to 37-kDa the combination may be useful in demonstrating the villus
extracellular glycosylphosphatidylinositol-linked glycoprotein smooth muscle (lymphatics) and the subepithelial myofibro-
that may also exist in a cleaved, soluble form. Thy-1 is blast network. Little is known of the peptides recognized by

Fig. 4. Identification of the intestinal mesenchymal cells. AC: confocal microscopic analysis of multicolor immunofluorescent staining of cross sections of a
normal human colonic crypt. CD90, ER-TR7, or TE-7 (red) identify nonhematopoietic mesenchymal cells of the colonic mucosa: fibroblasts, myofibroblasts, and
pericytes of the small vessels and lymphatics. The subepithelial myofibroblast network and pericytes are identified on the basis of their morphology, subepithelial
or perivascular location, and immunoreactivity to -SMA (green) and mesenchymal markers CD90, ER-TR7, or TE-7 (red), resulting in orange-yellow color on
the merged images. A: cell nuclei (blue) stained with DAPI; -SMA shown in green; Thy-1 (CD90) shown in red. B: cell nuclei (blue) stained with DAPI; -SMA
shown in green; ER-TR7 shown in red. C: cell nuclei (blue) stained with DAPI; -SMA shown in green; TE-7 shown in red. Calibration bars are 20 m.

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MYOFIBROBLASTS AND STEM CELL THERAPY
these antibodies other than their cytoplasmic intracellular lo-
cation.
Melanoma chondroitin sulfate proteoglycan. MCSP, also
known as neuron-glia 2 (NG2) in the rat, is a transmembrane
chondroitin sulfate proteoglycan expressed by immature pro-
genitor cells of the oligodendrocyte, chondrocyte, and smooth
muscle lineages (111). In the adult intestine, MCSP expression
is observed within myofibroblasts, pericytes, and smooth mus-
cle cells of the muscularis mucosae and tunica muscularis in
both the large and small intestine. MCSP and -SMA colocal-
ize within intestinal tissues (119). MCSP is not expressed by
fibroblasts and is rarely found in cells expressing fibroblast-
specific protein 1 (FSP-1; see Fibroblast-specific protein-1
below) (114). MCSP functions as a coreceptor for PDGF-AA,
basic (b)FGF, and integrins, effectively decreasing the thresh-
old dose for these ligands in MCSP-expressing cells (37). The
PDGF -receptor is unresponsive to PDGF-AA in smooth
muscle cells derived from the MCSP/NG2-null mouse (38).
MCSP expression is often enhanced within tumors, and we
have observed increased MCSP expression in stromal myofi-
broblasts isolated from human colorectal cancers, relative to
normal colonic tissue (R. C. Mifflin, unpublished observation).
In an elegant series of experiments, Huang et al. (48) have
recently demonstrated that MCSP/NG2-negative stromal vas-
cular pericytes severely limit the ability to assemble functional
tumor vasculature. The MCSP/NG2-null mouse demonstrates
defects in both pericyte recruitment and pericyte/endothelial
interactions resulting in reduced establishment of new blood
vessels (discussed in Ref. 48).
Fibroblast-specific protein-1. Fsp-1, also referred to as S100
calcium binding protein A4 (S100A4), is an intracellular pro-
tein identified by Strutz et al. (113) to be expressed in renal
fibroblasts but not epithelium. Subsequent studies have dem-
onstrated Fsp-1 expression by intestinal lymphocytes and mac-
rophages and malignant colorectal cancer epithelial cells (116,
118), but a fibroblast-specific transcriptional element has been
identified (87) that has been used in a series of elegant studies
to target transgene expression within fibroblasts (11, 88). Fsp-1
appears to be a useful marker to aid in the identification of
fibroblasts but is expressed at low levels, if at all, in myofi-
broblasts, pericytes, muscularis mucosae, and smooth muscle
of the lamina propria (116, 118). Sugimoto et al. (114) have
pointed out that within tumor specimens Fsp-1 antibodies
identify a unique population of fibroblasts with minimal over-
lap in expression of -SMA, MCSP, and PDGFR-. Thus
while Fsp-1 is useful for characterization and identification of
fibroblast populations, it is of little use as a myofibroblast
marker.
Patched. The receptors for hedgehog (Hh) growth and dif-
ferentiation factors are 12-span transmembrane receptors of the
patched (Ptch) family (125). Vertebrates contain two Ptch
genes, Ptch 1 and Ptch 2. Ptch normally interacts in an
inhibitory fashion with another transmembrane receptor,
smoothened (Smo), and Hh binding to Ptch disrupts this
interaction. Active Smo initiates a signaling cascade culminat-
ing in the activation and nuclear translocation of the transcrip-
tion factors Gli1, Gli2, and Gli3. Ptch 1 and Gli1 are them-
selves transcriptional targets of Gli transcription factors. In the
intestine of adult mice and humans, Ptch 1 (and Gli1) expres-
sion occurs predominantly within the myofibroblasts, peri-
cytes, and smooth muscle of the lamina propria and muscularis
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G689
mucosae. Hh signaling is crucial for the proper development of
myofibroblasts, muscularis mucosae smooth muscle, and lam-
ina propria smooth muscle since mice engineered to contain
low levels of intestinal Hh signaling contain significantly lower
numbers of these cell types. In these mutant Hh mice, the
epithelial compartment is deranged, displaying an increased
fraction of proliferating cells (65, 75, 132). These observations
underscore the importance of lamina propria and muscularis
mucosae -SMA cells to proper epithelial differentiation.
Periostin. Periostin is an ECM protein enriched in collagen-
containing structures such as ligaments, fascia, and cardiac
valves. Periostin is important for proper assembly of ECM
components in tissues exposed to high amounts of mechanical
stress (40). Periostin aids in the incorporation of tenascin-C
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into the ECM to generate a meshwork architecture of the
matrix (58). While a great deal has been published on the
functions of periostin in other tissues, little is known of its
expression and function within the intestine. Kikuchi et al. (59)
noted the synthesis and secretion of periostin by pericryptal
myofibroblasts and demonstrated a tight relationship between
-SMA and periostin expression in normal colonic mucosa.
The concordance between -SMA and periostin expression
diminishes in actively inflamed regions of ulcerative colitis,
where periostin is observed throughout the lamina propria and
-SMA expression remains in the pericryptal region and is
decreased (59). Much like what occurs in other epithelial
cancers (81, 103, 115, 120, 134, 135), periostin expression was
observed within the cancer-associated myofibroblasts of ad-
vanced colorectal adenocarcinoma (59). Periostin-expressing
fibroblasts, when cocultured with HCT116 epithelial cancer
cells in a three-dimensional collagen matrix, promote an in-
crease in the number and size of HCT116 colonies (59). Thus
while periostin expression represents an additional mechanism
whereby cancer-associated myofibroblasts promote tumorigen-
esis, periostin is a suitable marker for intestinal myofibroblasts
only under limited circumstances.
Fibroblast activation protein. Fibroblast activation protein
(FAP) is a type II transmembrane glycoprotein with post-
proline dipeptidyl aminopeptidase activity (4). FAP is highly
expressed by myofibroblasts and pericytes within the stroma of
numerous cancers (106, 129), including colorectal adenocarci-
noma (127), but is generally undetectable within normal tissues
(104, 106). FAP-deficient mice display retarded growth of lung
and colon tumors relative to the parental strain, and pharma-
cological or immunologic inhibition of FAP activity also leads
to decreased tumor growth in animal models (19, 45, 70, 106,
127). FAP depletion retards tumorigenesis by causing elevated
collagen accumulation, decreased myofibroblast content, and
decreased blood vessel density (106). FAP expression has also
been observed in myofibroblasts within tissues undergoing
fibrotic episodes such as idiopathic pulmonary fibrosis, hepatic
cirrhosis, and rheumatoid arthritis and within strictures result-
ing from Crohns disease (104). Bae et al. (8) have determined
that MSCs derived from bone marrow, adipose tissue, and
umbilical cord blood constitutively express FAP. Immunose-
lection using a FAP monoclonal antibody yielded a homoge-
neous population of MSCs (8). Thus while FAP can be used as
a marker of activated myofibroblasts and pericytes within
tumors and fibrotic tissues, it may also be used for enrichment
of their immediate precursors, MSCs.
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G690 MYOFIBROBLASTS AND STEM CELL THERAPY
Epimorphin. Epimorphin/syntaxin 2 is a membrane-associ-
ated protein that is homologous to the syntaxin family of
vesicle-docking proteins in neurons and pancreas, where they
take part in calcium-mediated exocytosis. Epimorphin levels
increase in the fetal gut mesenchyme during lumen formation
and villous morphogenesis (32). In the intestine, epimorphin is
predominantly expressed by subepithelial myofibroblasts and
vascular pericytes (6), although expression by other stromal
cell types cannot be ruled out (109). Epimorphin regulates the
cellular response to Hh ligands, and myofibroblasts isolated
from Epi/ mice have a reduced response to Hh, and thus
decreased Bmp signaling (109). Myofibroblasts engineered to
overexpress epimorphin secrete increased levels of Bmp4,
which promotes epithelial differentiation.
Origin of Lamina Propria Mesenchymal Cells
Newly proposed therapy for severe and nonresponsive IBD,
especially Crohns disease, is bone marrow transplantation or
stem cell therapy (see Targeting Subepithelial Mesenchymal
Cells for Stem Cell Therapy below). There are many mecha-
nisms by which stem cell therapy might be effective for
autoimmune diseases (also see below): replacement of diseased
pericryptal myofibroblasts, which form the epithelial stem cell
niche and also regulate epithelial cell differentiation; the re-
placement of diseased pericytes and endothelial cells, which
are necessary for angiogenesis; and even engraftment and
transformation of stem cells into neoepithelial cells are theo-
retically possible. Effective therapy by stem cells requires
knowledge of which bone marrow stem cell, mesenchymal
(MSC) or hematopoietic (HSC), is critical for the normal stem
cell repair process in the intestine as well as how these cells
induce repair.
Origin in the embryo. In the human intestine, lamina propria
-SMA mesenchymal cells are present in the embryo as early
as 21 wk, and probably earlier (107), and in the mouse they
appear at embryonic day 18.5 (80). These cells have the
appearance of myofibroblasts. Proper development of subepi-
thelial mesenchymal cells is dependent on epithelium-derived
Hh signaling (64, 75, 125, 132). Proper localization of lamina
propria mesenchymal cells during development requires
PDGF-A and PDGFR-, while PDGF-BB and PDGFR- are
needed for proper pericyte formation in the kidney and skin
(the intestine was not well examined) (52, 73). There is
controversy (reviewed in Ref. 99) over whether these subepi-
thelial mesenchymal cells originate from neural crest cells,
which has been demonstrated for mesenchyme and pericytes of
the thymus (31, 82, 131), or from the serosal mesothelium
(128).
Subepithelial mesenchymal cell proliferation is driven by
PDGF-BB, EGF, bFGF, and IGF-I and -II and better still in a
combinatorial signaling process achieved by pairing PDGF
with IL-1 or vasoactive polypeptide (VIP) (49). Combina-
tions of TGF- and EGF or IGF-II may achieve significant
fibroblast proliferation through a connective tissue growth
factor (CTGF)-dependent pathway (39).
Origin in the adult. Gabbiani (46), one of the early pioneers
of myofibroblast biology, proposes that other mesenchymal
cells in organs such as -SMA fibroblasts and -SMA
stellate cells, pericytes, and even smooth muscle cells might
become myofibroblasts in the postnatal animal through pro-
AJP-Gastrointest Liver Physiol VOL
cesses of differentiation, activation, and dedifferentiation (re-
viewed also in Ref. 99). Furthermore, both epithelium and
endothelium might transdifferentiate to myofibroblasts through
the process of epithelial to mesenchymal transition (EMT) (95,
133). Finally, subepithelial -SMA cells may be replenished
in the postnatal animal by stem cells from the bone marrow.
Brittan et al. (14) described pericryptal myofibroblast replace-
ment in female recipients by male bone marrow transplantation
through identification of the Y chromosome. This group later
demonstrated multiple organ engraftment after bone marrow
transplantation (26) and neoplastic recruitment of transplanted
bone marrow cells (15). Engraftment of transplanted cells into
tissue myofibroblasts can be increased by wounding the
tissue epithelium, and the percentage of donor-derived colonic
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myofibroblasts and vascular pericytes can reach 39% within a
week of transplantation following trinitrobenzenesulfonic acid
(TNBS) colitis (13). A similar phenomenon occurs after dam-
age to lung, kidney, stomach and skin, where engrafted mes-
enchymal cells can occupy 42%, 24%, 64%, and 4% of
resident subepithelial mesenchymal cells, respectively, within
10 wk of transplantation (26). Thus the role for bone marrow
stem cell replacement of damaged intestinal stromal cells is
clear, but these experiments raise two important questions:
What is the nature of the stem cell aiding in tissue repair? Do
the stem cells only replace lamina propria mesenchymal cells
or do bone marrow stem cells also replace damaged epithelial
cells?
Targeting Subepithelial Mesenchymal Cells for Stem Cell
Therapy
Bone marrow transplantation and stem cell therapy have
been proposed as a new therapy for severe and nonresponsive
IBD, especially Crohns disease. The rationale behind such
therapy is multiconceptual (Table 1). HSC transplantation
(HSCT) can accomplish immune reset, i.e., the generation of
new self-tolerant lymphocytes after chemotherapy-induced ab-
lation or reduction (conditioning chemotherapy) of self-reac-
tive lymphocytes (33). In the case of allogeneic (donor) trans-
plants, the replacement of genetically abnormal bone marrow
precursors with healthy HSCs is also possible. The idea that
MSC therapy might topically engraft and be used to replace
diseased subepithelial mesenchymal cells with healthy trans-
Table 1. Therapeutic effects of stem cell therapy in IBD
Hematopoietic stem cell transplantation
Immune reset: generation of new self-tolerant lymphocytes after bone
marrow ablation or conditioning chemotherapy
Replacement of genetically abnormal bone marrow precursors through
allogeneic transplantation
Stromal cell engraftment in the lamina propria with healthy cells,
increasing growth factor production, which creates a healthy stem cell
niche and promotes angiogenesis/vasculogenesis
Possibly engraftment followed by EMT to replenish epithelial cells
Mesenchymal stem cell transplantation
Immune suppression: MSCs are immunoprivileged cells
Stromal cell engraftment in the lamina propria with healthy cells,
increasing growth factor production, which creates a healthy stem cell
niche and promotes angiogenesis/vasculogenesis
Possibly engraftment of MSCs followed by EMT to replenish epithelial
cells
IBD, inflammatory bowel disease; EMT, epithelial to mesenchymal transi-
tion; MSC, mesenchymal stem cell.
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MYOFIBROBLASTS AND STEM CELL THERAPY
planted stromal cells, or that stem cells might replace and
transdifferentiate into epithelial cells, is a distinct possibility
(69). Alternatively, because MSCs demonstrate significant im-
munosuppressive properties when infused in vivo (see Mesen-
chymal stem cells below), this property may be used as a
mechanism of therapeutic response to MSC therapy in auto-
immune diseases. Effective therapy with stem cells requires
knowledge of which bone marrow stem cell, mesenchymal
(MSC) or hematopoietic (HSC), is most critical for the normal
stem cell repair process in intestine and the mechanism of that
response.
Hematopoietic stem cells or fibrocytes. HSCs express a set
of characteristic molecules: the leukocyte common antigen
CD45 (pan-hematopoietic marker), progenitor markers CD34
and CD105, monocyte markers such as CD11, CD13, and
CD14, and antigen-presenting molecules such as MHC class II
and the T lymphocyte B7 family costimulatory molecules
CD80, CD86, and also CD40. Like MSCs, they express vi-
mentin, chemokines and chemokine receptors, adhesion mol-
ecules, and integrins. They do not normally express MSC
markers such as CD90, CD44, CD70, CD73, and CD271.
A recent review details current therapeutic outcomes with
myeloablative (chemotherapeutic ablation of both the hemato-
poietic and lymphopoietic lineages in the bone marrow) HSCT
performed for treatment of cancer in patients with concomitant
IBD (25 patients in 10 separate reports). Described also are the
outcomes of the transplantation of 19 Crohns disease patients
in 5 reports who underwent nonmyeloablative, conditioning
chemotherapy, which depletes self-reactive lymphocytes. Clin-
ical remissions were noted in all and lasted at least 6 mo and
up to 7 yr (35). The benefits of these transplantations were
thought to be due to the immune reset that occurs with
hematopoietic cell replacement (Table 1). Is there experimental
evidence of intestinal mesenchymal or epithelial engraftment
after HSC transplantation?
Using carefully performed, single-cell cloning of green flu-
orescent protein (GFP)-positive HSCs, Ogawa et al. (85) have
demonstrated that HSCs can repopulate pericytes in the kidney
(mesangial cells), brain, and heart. While it is possible that the
HSC itself populated these distal sites (78, 110), it is likely that
tissue engraftment occurs through an intermediate cell, a he-
matopoietically derived monocyte derivative called the fibro-
cyte (reviewed in Refs. 9 and 20).
Fibrocytes are a distinct population of bone marrow-derived
cells that become progenitors for mesenchymal cells. They
express hematopoietic markers of a monocyte origin and also
produce fibroblast/myofibroblast ECM molecules and matrix-
modifying enzymes such as matrix metalloproteinases (MMPs)
and tissue inhibitors of metalloproteinases (TIMPs) (9) (Fig. 5). In
the vascular compartment, where they are estimated to repre-
sent 0.1 0.5% of circulating cells, they express CD45, CD34,
CD105, CD11, MHC class II, CD80 and CD86, and CD13, as
do HSCs, but are only weakly positive for the monocyte
marker CD14 and negative for CD16. They are also negative
for B-cell and T-cell markers and do not express dendritic
markers CD1a, CD10, or CD83. They express chemokine
receptors CCR1, 3, 4, 5, 7, and 9 as well as CXCR1, 3, and 4.
Importantly, they express fibrocyte/mesenchymal markers vi-
mentin, collagen 1, fibronectin, and MMP-9. It is unclear
whether they express the fibroblast marker CD90 (Thy-1) (9).
In the human it is thought that they are derived from a small
AJP-Gastrointest Liver Physiol VOL
Review
G691
Downloaded from http://ajpgi.physiology.org/ by 10.220.33.2 on September 13, 2017
Fig. 5. Bone marrow stem cell origin of intestinal mesenchymal stromal cells.
Diagram demonstrates possible stem cell origin and routes for derivation of
lamina propria mesenchymal cells (and epithelial cells) from bone marrow
precursors. See text for discussion. TGF-, transforming growth factor-;
MCSP, melanoma chondroitin sulfate proteoglycan; PDGFR-, PDGF recep-
tor-.
subset of CD14, CD16 monocytes that bear surface chemo-
kine receptor CCR2 (the analogous murine counterpart would
be Ly-6Chigh, CRR2). These low-abundance cells belong to a
group of white blood cells termed inflammatory monocytes
that normally replenish tissue macrophages and dendritic cells.
In culture, and presumably in vivo, after direct contact with T
cells a subgroup of these cells becomes fibrocytes and there is
downregulation of CD14 and upregulation of collagen synthe-
sizing enzymes, MMPs, IL-1, and PDGF-AA (1). In inflamed
tissue where they are exposed to TGF- and ET-1, there is
upregulation of -SMA and downregulation of CD34 and
CD45 and the cells become myofibroblasts. These cells have
been incriminated in several pathological processes and dis-
eases including hypertrophic scar and keloid formation, airway
remodeling in asthma, interstitial pulmonary fibrosis, renal
fibrosis, pancreatic fibrosis, and even atherosclerosis (1, 9,
108).
Recently, Uehara et al. (123) have published evidence that
fibrocytes may be involved in colonic repair in colitis. Seven
days after the induction of dextran sodium sulfate (DSS)
colitis, there was an influx of CD45-positive, collagen 1-pos-
itive ovoid cells into the inflamed tissue. By day 14, -SMA-
positive spindle-shaped myofibroblasts were present in the
tissue. Unfortunately, the study does not definitively demon-
strate the proposed sequence of fibrocytes becoming myofibro-
blasts because the lack of cell labeling of the bone marrow
precursor cells does not allow lineage tracing and thus defin-
itive identification of the HSC origin of the fibrocytes in this
intriguing experiment.
Mesenchymal stem cells. The properties of MSCs in animal
and humans and their role in normal health and disease have
been reviewed elsewhere (17, 29, 122). MSCs are cultured
from bone marrow stromal cells by adhesion to plastic in 10%
calf serum. They can serve as feeder layers for the culture of
HSCs and, with proper conditions and the addition of proper
growth factors, can self renew and differentiate into mesoder-
300 MAY 2011 www.ajpgi.org
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G692 MYOFIBROBLASTS AND STEM CELL THERAPY
mal tissue such as adipocytes, chondrocytes, osteocytes, and
other tissue stromal cells. They also show multipotentiality by
transdifferentiating, under the influence of different factors,
into cells of other germ line lineages such as endoderm (lung
and intestinal epithelial cells and muscle cell) and ectoderm
(skin epithelial cells and neurons). Whether such transdiffer-
entiation occurs in vivo has been controversial, but this seems
likely under conditions of organ damage and repair (112).
MSCs express characteristic molecules such as CD90 (Thy-1),
CD44 (hyaluronan receptor), CD105 (SH2), CD73 (SH3/4),
CD90, CD71, CD271, and STRO1. Cultured MSCs are defined
also by their failure to express HSC markers such as CD45
(pan-hematopoietic marker), CD34 (stem cell progenitor
marker), CD14 (monocyte marker), CD11 (monocyte/dendritic
marker), and CD4 T-cell antigen-presenting molecules such as
MHC class II and the costimulatory molecules CD80, CD86,
and CD40. MSCs express several adhesion molecules such as
VCAM-1, activated leukocyte adhesion molecule (ALCAM),
ICAM-1, ICAM-3, endoglin (CD105, TGF- coreceptor) and
various 15- and 1,3,4-integrins as well as many of the CC,
CXC, CX3C, and XC chemokine receptors, which aid in their
ability to home to and adhere to damaged tissues. It is impor-
tant to note that in some animal species MSCs may not express
the same molecules as human MSCs, e.g., mouse MSCs may
show variable CD34 expression, and MSC expression patterns
in vivo may not be the same as in established culture. Although
typically cultured from bone marrow, isolation of MSCs has
been accomplished from fetal blood, umbilical cord, amniotic
fluid, and adipose tissue (17).
In addition to their characteristic ability to support HSC
growth and differentiation, and their ability to differentiate into
mesenchymal tissues, another fundamental property of MSCs
is their immunomodulatory properties, with effects on both
innate and adaptive immunity (reviewed in Refs. 29, 99, and
122). The MSC is known as the immunoprivileged cell or
universal donor. It is important to understand that all mature
mesenchymal tissue stromal cells share these immunoregula-
tory properties (50). Some investigators have proposed that
tissue MSCs might gain MHC class II molecules with the
ability to present antigens and promote inflammation (91) or
suppress inflammation through expression of PD-L1 immune
suppressor molecules, and this bivalent immune function might
be governed by the level of ambient IFN- (18). Most of the
salutary effects of infused MSCs in animal models of organ
transplant rejection, central nervous system (CNS), kidney,
heart, liver, and pancreas, are attributable to these immuno-
modulatory effects (29, 130). In other disease models of heart,
lung, and CNS the beneficial effects of transplanted MSCs
have been attributed to the trophic effects of MSC-derived
growth factors that induce proliferation and differentiation of
local, host progenitor cells and not to topical engraftment of
these MSCs with subsequent incorporation into the tissue (17,
35, 69, 122). Recently MSCs have been used to treat graft
versus host disease, type 1 diabetes, and Crohns disease, but it
is too early to determine therapeutic efficacy.
There have only been a few studies of the ability of MSCs
to engraft and repair colitis. Brown et al. (16) in a model of
DSS colitis recognized a COX-2-expressing stromal cell that
moved from the upper regions of the colonic lamina propria to
the basal regions of the lamina propria adjacent to the location
of epithelial stem cells. This repositioning was dependent on
AJP-Gastrointest Liver Physiol VOL
MYD88 signaling, indicating that toll-like receptors were im-
portant to the directional movement of this prostaglandin-
secreting cell. Furthermore, this study showed that prostaglan-
dins were critical in initiating epithelial proliferation and re-
pair. Unfortunately, there was no confirmation of the origin of
this cell: it could be an MSC or a tissue fibrocyte.
In mice (57) or rats (117) with DSS-induced colitis or in rats
with TNBS-induced colitis (43), infusion of MSCs into nonmy-
eloablated animals improved the severity and shortened the
duration of colitis. The MSCs were topically implanted into
the lamina propria. The beneficial effects were ascribed to
the provision of growth factors and angiogenic/vasculogenic
factors or differentiation into endothelial cells and other
lamina propria mesenchymal cells (fibroblast, myofibro-
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blasts, and smooth muscle cells).
There is evidence that transplanted bone marrow stem cells
may engraft topically into the gut epithelium as well as into the
lamina propria. With Y chromosome identification, bone mar-
row-derived cells can be found in the intestinal epithelium of
humans (79, 89). In experimental animal models, damage to
the epithelium is thought to create chemokines that enhance
homing of stem cells, and when GFP-labeled bone marrow
cells were transplanted into a TNBS mouse model of colitis,
donor cells were found in 38% of colonic epithelial cells after
transplantation (60). While the lineage of these engrafting cells
is uncertain, Yabana et al. (130) used male donor MSCs
derived from GFP engineered rats and infused them into
wild-type, DSS-colitis female mice either with normal marrow
or with busulfan myeloablation. The transplanted cells were
identified with colocalization of eGFP and Y-FISH analysis.
Transplanted MSCs were found in the lamina propria adjacent
to the crypts (presumably subepithelial myofibroblasts or peri-
cytes); however, the bulk of the cells were found engrafted into
the epithelium: 2% of colonic crypts and 10% of colonic
epithelial cells bore the GFP marker in the group with normal
marrow and 8% of crypts and 42% of epithelial cells in the
myeloablated model (130). These investigators also demon-
strated improved barrier function in the MSC transplanted
animals. Whether there is direct engraftment of bone marrow
stem cells into the epithelium after infusion of MSCs or
whether EMT of engrafted subepithelial myofibroblasts oc-
curred is uncertain.
Mesenchymal cells of fibrotic lesions. A major consequence
of the chronic inflammation and tissue injury associated with
IBD, particularly Crohns disease, is tissue fibrosis (102).
Fibrosis results from excessive local accumulation of ECM
molecules laid down by activated myofibroblasts and fibro-
blasts or by resident stellate cells. These activated stromal cells
may be derived locally via stimulation or induction of prolif-
eration, or by EMT driven by proinflammatory cytokines
(102). Additionally, resident myofibroblasts and fibroblasts
may migrate to sites of inflammation in response to chemot-
actic gradients of PDGFs, IGF-I, or TGF-. As mentioned
above, stromal cells derived from MSCs, fibrocytes, or HSCs
may also home to sites of inflammation and, in the presence of
ongoing inflammation, become activated myofibroblasts/fibro-
blasts and contribute to tissue fibrosis.
The key to therapy directed at preventing fibrosis in IBD is
to eliminate or curb the chronic inflammation. The immuno-
modulatory capacity of MSCs and their ability to provide
angiogenic/vasculogenic factors and trophic factors for re-
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MYOFIBROBLASTS AND STEM CELL THERAPY
placement of damaged epithelium suggest that MSC transplan-
tation may play a useful role in the treatment of IBD-related
intestinal fibrosis. Additionally, it is possible that mesenchymal
cells newly arrived as stem cells and then integrated into the
lamina propria might be less profibrogenic than those present
during chronic inflammation. While the exact mechanism is
unclear, preliminary studies using animal models of hepatic,
lung, and cardiac fibrosis support the conclusion that stem cell
therapy may be useful antifibrotic treatment (28, 41, 71, 72).
Conclusions
Mesenchymal cells of the intestinal lamina propria represent
a spectrum of phenotypes that perform distinct functions based
upon their location. The first layer of subepithelial cells sur-
rounding the crypts of the colon and small intestine are appar-
ently myofibroblasts, although no doubt pericytes are present
in the lamina propria also. In the surface of the colon and villus
of the small intestine, the subepithelial cells are predominantly
pericytes. Additional pericytes are found surrounding blood
vessels and lymphatics in the core of the villus, and these cells
seem to be connected to subepithelial cells as part of a
three-dimensional network. The second layer of mesenchymal
cells surrounding crypts, the mesenchymal cells of the villus,
and the subsurface areas of the colon, the interconnecting
mesenchymal cells of the villus core cells lose -SMA staining
and become fibroblasts. Lymphatic smooth muscle and peri-
cytes are also present in the lamina propria.
Immunologic markers exist to aid in the identification of
these cells, but none is absolutely specific or definitive. Lack of
desmin expression appears to help separate -SMA myofi-
broblasts from pericytes (weakly positive for desmin) and from
the smooth muscle of the muscularis mucosae and lymphatic
lacteal (strongly positive for desmin). Vimentin staining is less
intense or absent in these smooth muscle bundles. Coupling
-SMA staining with antibodies marking thymus stromal cell
antigens appears to be useful in separating myofibroblasts and
pericytes from fibroblasts.
The primary origin of lamina propria mesenchymal cells in
the embryo remains uncertain. In the postnatal animal subep-
ithelial mesenchymal cells (myofibroblasts and pericytes) can
be recruited from circulating precursor pools during inflamma-
tion and carcinogenesis, but it is unclear whether the important
pool is originally from an HSC origin, from a MSC origin, or
from both. Subtle functional differences may exist among cells
derived from either MSCs or HSCs, but such differences have
yet to be identified.
MSCs and HSCs represent future therapeutic hopes for
intestinal injury and chronic intestinal inflammatory states.
Current stem cell therapies are directed toward the immune
aspect of intestinal inflammatory diseases. However, studies in
animal models suggest that stem cell therapies might replace
lost stromal cells (and perhaps epithelial cells) after injury.
Thus an important aspect of stem cell therapeutic potential may
represent true regenerative medicine and be derived from their
replenishment of the subepithelial mesenchymal cells that
normally play important roles in growth, development, and
repair of the intestinal mucosa.
GRANTS
The research performed in the authors laboratories mentioned in this article
was supported by National Institutes of Health Grants DK-55783 (D. W.
AJP-Gastrointest Liver Physiol VOL
Review
G693
Powell) and CA-127229 (D. W. Powell) and an American Gastroenterological
Association (AGA) Scholars Award to I. V. Pinchuk.
DISCLOSURES
No conflicts of interest, financial or otherwise, are declared by the author(s).
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