PREPARATION OF SERUM AND CHICK EMBRYO EXTRACT IN ANIMAL CELL CULTURE

PRESENTED BY: MD.WASEEM GUIDED BY: MR. S. KSHIRSAGAR

Steps Involved In Serum Preparation

    

Collection Of Blood Clotting Separation Of Serum Sterilization Storage

1.Collection Of Blood
(Depending upon the Volume required)
For Large Volumes
Arrangements are made at the Slaughterhouse and Blood is collected directly from the bleeding carcass.

For Small Volumes

Blood withdrawn directly from live animal under veterinary supervision.

2. Clotting


Natural Clot Serum

Allow the blood to clot having it stand overnight in covered container at 4C.

Physical Separation
Centrifugation of blood followed by defibrination.

Natural Clot Serum is superior to Physically Separated Serum

3. Separation Of Serum


Separate the Supernatant

Centrifuge for 1Hr to remove the sediment

4. Sterilization


Serum Is Sterilized By Filtration through 0.1m porosity. -Due to its viscosity and high particulate content,
the serum should be passed through series of fiberglass or other prefilters before passing through the final sterilizing filter.

5. Storage


 

 

Bottle the Serum in sizes that will be used up within two or three weeks after thawing. Freeze the serum as rapidly as possible It is best used within 6-12 months of preparation if stored at -20C. For more prolonged storage, store at -70C. Polycarbonate or High density polypropylene bottles will eliminate the risk of breakage at these temperatures.

Preparation of Chick Embryo Extract

1.

2. 3.

4.

Remove embryo from egg and place it in Petri dish Take out its eyes Transfer it to flat or round bottomed 50ml containers. Two embryos in each container Add equal volume of Hank s BSS

Preparation of Chick Embryo Extract

5.

6.

7. 8. 9.

Mash the embryos in the BSS with sterile Glass rod previously heated and flattened at one end until they broken up Let the mixture stand for 30min at room temperature Centrifuge for 15 min Remove supernatant Store at -20C

Preparation of Chick Embryo Extract

Extract of Chick and other tissue may also be prepared by Homogenization in a Potter Homogenizer or Waring Blender

Preparation of Chick Embryo Extract by Homogenization

1.

2.

3.

4. 5.

Homogenize chopped embryos with an equal volume of Hank s BSS Transfer homogenate to centrifuge tubes and centrifuge for 10 min Collect supernatant n again centrifuge it for 20min Check sample for sterility Store at -20C