hTERT method for immortalization - Anyone with any experiences? (Sep/21/2006 )
ATCC promotes immortalization with human telomerase reverse transcriptase (hTERT) by introducing a hTERT gene
as I understand if transfected (or infected), the gene product hTERT "immortalizes" the cell; so, one has to select a continuously hTERT-expressing clone, and gets an immortalized cell line; sounds quite simple but has anyone experience with hTERT method of immortalization of f.i. primary cells?
(this question was asked in a reply some days ago, and is here asked again to appeal more by headline and own topic)
I've not got personal experience of this but in my old department they did this with primary fibroblasts. They did in collaboration with a pathology group who work on telomerase. I think they used a retorvirus expressing hTERT and I think the fibroblasts no longer became senenescent after 30 passages because of the telomerase activity. The reference is below.
All the best,
McSharry BP, Jones CJ, Skinner JW, Kipling D, Wilkinson GW.
Human telomerase reverse transcriptase-immortalized MRC-5 and HCA2 human fibroblasts are fully permissive for human cytomegalovirus.
J Gen Virol. 2001 Apr;82(Pt 4):855-63.
thanks for competent reply and your good wishes!
Have you immortalized the cell?
in the near future wel like to try