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immortal or stable cells - primary human cells to immortal cell line? (Jun/24/2009 )

I work with primary cells from fetal membranes and having difficulty coordinating complicated protocols and being dependent on collection of cells from a clinical source. We are interested in trying to immortalize these epithelial cells but I cannot find any protocols to accomplish this. Can anyone offer any suggestions?

-murth002-

Transfect with large T antigen from adenovirus, will still give only about 1-5% of cultures transformed, but should work OK otherwise.

Be aware that transformed cells are not at all similar to primary cells.

-bob1-

bob1 on Jun 24 2009, 04:20 PM said:

Transfect with large T antigen from adenovirus, will still give only about 1-5% of cultures transformed, but should work OK otherwise.

Be aware that transformed cells are not at all similar to primary cells.


Hi!!!

Don't know if it will work well for you but Epstein-Barr virus, a member of Herpesvirus family is able to immortalize lymphocites. Other virus are Human papillomavirus (HPV) a member of Papilomaviridae family and Simian Vacuolating virus 40 (SV-40), a member of Polyomaviridae family (monkeys).
Unfortunatelly i don't have any protocol to immortalize these cells line or others even i have never done it before, but i think so it will be easy to get one in Pubmed.

Regards...

Reis,V.P.

-Cell buster-

Yeah, other viruses will work too. I gave the T-antigen (oops, I was meaning SV-40 T-antigen, adenovirus E1b55K) one as it is possible to use a plasmid rather than having to infect with viruses and all the biosafety issues related to them.

-bob1-