cell proliferation assay

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contingency
contingency's picture
cell proliferation assay

hi all,
resently, i use MEF cells to do a cell proliferation assay with bFGF, but i found that when i add it into the culture it can`t promote cell proliferation compare to the control group and those cells treated with bFGF really have a morphological change at the same time. so i suspect the bFGF concentration i used in my experiment. can any one here give me some advices?
thanks.

Omai
Omai's picture
Hi contingency,

Hi contingency,

Have you tried titering the concentration of bFGF. What concentration are you using now? Also, what measure of proliferation are you using?

I have found that growth factors like bFGF are very sensitive to multiple freeze thaw cycles and can degrade very quickly. Are you using a fresh stok of bFGF?

Omai

contingency
contingency's picture
hi Omai,

hi Omai,
My bFGF concentration used in the experiment  is 50ng in one milliliter DMEM with 10%FBS and i add it in to medium every two days. is that too high? i saw 5 or 10ng/ml used in some papers. it stocked at -80 degree before use. and i just use a haemacytometer to count cell number.
thank you for your attention. could you give me more advice about this?

contingency