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?
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 on Jun 24 2009, 04:20 PM said:
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.
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.