I'm going to start culturing human immortalized podocytes and I want some tips about the conditions of culture media. My goal is to perform treatments with different concentrations of glucose and I have some doubts about the composition of media in the distinct phases. I have been reading a lot of articles, but there is not a clear explanation. For example, for proliferation phase its clear that I have to use RPMI media, 10% FBS, ITS and antibiotics. But for differentiation phase: I should use a media with only 2% FBS supplement (or another percentage), and without ITS? I have read that FBS reduction helps differentiation, but also I have read that induces apoptosis.

And in a pre-treatment phase: there is a serum-glucose stravation?And without ITS too? Or supplement media with 5mM glucose? For how many time?

And finally, in the treatment phase, I don't know if ITS is necessary too, and if serum percentage is also low (1-2%).

Please, if someone could help me I would appreciate it.

Thank you!