I have irradiated MEF cells which will be frozen to be used to produce feeder layers for the growth of iPS cells. I have seen two methods from various protocols; seems I should either use MEF media (with 10% FBS and 10% DMSO) or simply use neat FBS (with 10% DMSO). Is there an advantage or disadvantage to either formulation?
Hi Sophie, I would go with 90% FBS and 10% DMSO and do a slow freeze at -80oC. Wrap up the cryovials containing your cells like a mummy and put them in the -80oC freezer. After ON, put them in liquid N2.
Just be sure that the DMSO is tissue culture grade. I haven't lost a cell line yet using this procedure.
Hi Sophie,
I am using 1/2 MEF medium (10%FCS) +1/2 80% FCS/20% DMSO which equal to 45%FCS/10%DMSO. This way you use less serum (which can be really expensive) and my irradiated MEF are fine.
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