I am trying to grow non-differentiated PC12 cells in RPMI 1640 10%FBS+5%HS, passing every third day. Recently I have realised that many of cells are dead.
I have thawed two aliquots frozen at different time points/passage numbers. And now I see that in all three flasks cells die.
I have just checked them by flow cytometry - only ~4% is alive.
Any ideas, why?
Hi! According to my experience, PC12s usually grow well when they are adhered on the plates. Unless your culturing condition is necessary to be suspension, culturing PC12s at steady condition, to let them adhere on the plate, would help. Additionally, I used different composition of media, 10% FBS+10% HS (horse serum).
It looks like the problem was the medium. Unfortunately I didn't have another bottle of the same one, and it took time for it to be delivered. So I took another medium I had (DMEM+15%FBS), and PC12 was quite happy to grow on it.
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