Serum Starvation - Troubles with Serum Starvation of NIH 3T3 (Mar/12/2005 )
In our lab we tried to synchronize NIH 3T3 cell line by a 18 hour-serum deprivation. The result of our starvation was a 75-80% of cell death after the 18 hours.
Can anybody tell me which is the right way to starvate this cell line? Thanks a lot for your attention!
That's strange, NIH-3T3 cells can even grow if you throw them on the floor and let them there! When I did cell division experiments culturing the cells with several hormones and factors, my controls in serum-free media didn't die after a 5-days incubation, so 18h incubation shouldn't kill them. Which is the recipe of your serum-free media?
I have the same problem, my cells also de-attach after about 48 hours of serum deprivation. I use DMEM with glutamine and antibiotics. Need I use something else?
Thank you very much
have you diminished the concentration of Atb?
i know that Atb bind to serum proteins and then are less toxic in 10% FBS-suppl medium...
I have the same troubles with 3T3.
There is a paper that serum starvation induces apoprosis of NIH3T3 (Chen HH, Zhao S, Song JG. TGF-beta1 suppresses apoptosis via differential regulation of MAP kinases and ceramide production. Cell Death Differ. 2003 May;10(5):516-27.)
hi all ! serum deprivaton should be done in a gradient fashion so that the cells do not enter in to a state of shock. gradually decrease the FBS concentration in two or three steps. minimum being 2.0 % to 0.5%. and deprivation should be done not for not more than 16-18 Hrs. of course it depends on the cell type. but for most cell types its 16-18 hrs. apoptosis sets in thereafter.
all the best