Hello everybody,

I am trying to synchronize MEFs (Mouse Embryonic Fibroblasts) in the S phase, but I have had no success. I have already tried a synchronization with serum starvation (24 h of starvation+ 18h of release), double thymidine block (for some reason had no effect on synchronization, flow cytometry showed just a normal cell cycle), and treatment with hydroxyurea (have tried 0.5 mM, 1mM, 2 mM and 4 mM concentration for 20 h and all of them just kill the cells)

Does anyone have expertise in the synchronization of MEF cells, so you can give me some advice?

Thanks in advance