HI everyone! I am going to start culturing RPTEC/hTERT renal proximal epitelial cells and I have several questions about it. First of all I want to know when the cells are proliferating and when differentiating. I have cultured immortalized podocytes previously and these cells proliferate at 33ºC and differentiate at 37ºC, so i have clearly differentiated the two populations. But in the case of RPTEC/htert cells I dont know how to know it, because I think they dont have the SV40 T antigen to thermoswitch them. So, are cells proliferating and differentaiting joint? How to know which are mature cells? And if I want to treat them with something, I will treat proliferating and differentiating cells?
Another question is the appropiate culture medium for these cells. I have read that the supplied medium from evercyte or ATCC in some cases didn't work very well, and as well the mixture of DMEM/HAMs F12 . So the best option is to buy DMEM (low glucose with glutamine?) and F12 separately, plus glutamax, EGF, ITS, Penicillim/Streptomycin and hydrocortisone? and FBS is necessary or not?? Because I have read the two options
I would appreciate any answer from peolple working with this cell line.
Thank you in advance
Ana
RPTECs that are immortalized by hTERT may not exhibit differentiation markers (see link below) in culture.
Immortalized cells tend to adapt culture conditions. Therefore, some optimization of culture conditions (including the culture medium, growth factors, and culture container source) may help you decide the optimum conditions.
Article Differentiation Status of Human Renal Proximal and Distal Tu...
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