I am trying to culture murine cell line (lymphocytes) in medium supplemented with serum of mouse bought from Sigma (cat# M5905). I inactivated the serum in water bath at 56°C for 1hour. Next I made some tests with differents concentrations of mouse serum (5%, 10% and 20%), but the cells died before 24h. The controls (without serum and with 10% FBS) did not die in 24h.
Someone knows what is happening or if there is other way to inactivate the serum?
Is it possible that the 56C actually inactivated cytokines that maybe very important for lymphocyte to survive?
As stated, you should follow supplier recommendations. You can also try with supplements of Glutamine and non essential aminoacids. Some cell lines need these supplements but can grow under FCS or FBS without them, though a slower rate. Perhaps your cell line is one of those, and when shifting to mouse serum they need the supplements.
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