How to avoid contamination of primary cells during preparation?

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How to avoid contamination of primary cells during preparation?..

Answer / anil kumar pr

The entire procedure of primary cell isolation and culture
should be carried out under strict aseptic condition as in
any cell culture practice. However, to minimize the chances
of contamination arising from the cell source such as
animal, organs or tissues, certain pretreatments are necessary.
1. As soon as the tissue is collected, place the it in PBS
(or any suitable buffer) containing high concentration
antibiotics (10X penicillin and streptomycin) for 2-5 min.
2. Immediately transfer the tissue to biosafety cabinet.
3. Rinse with buffer containing antibiotic (1X) (1-2 times).
4. Rinse with sterile PBS (3 - 5 times).
5. Transfer the tissue to buffer in a sterile vial and
remove all other items used for pretreatment from the
biosafety cabinet; such as waste buffer, tissue bits, used
pipette tips, container for waste collection etc.
6. Wipe the surface of BSC with 70% alcohol.
7. Wait for 2 minutes with 'flow' switched on.
8. Start cell isolation procedure.

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How to avoid contamination of primary cells during preparation?..

Answer / n.c.pandey

Use of sterile wares,medium and use of Laminar flow hood
while preparing the primary cells.

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