I'm trying to differentiate C2C12 cells using various media. I've tried 1% and 2% of horse serum and also insulin. The cells that received only HS did not differentiate (many died after 24h, the remaining changed the shape), the ones with insulin differentiated pretty well. Plates 100% confluent and with proliferation medium also differentiated spontaneously.
I know that the standard protocol uses only HS. So why are not my cells able to differentiate?
Hello
I'll give you some notes , i hope help you
1.) propagate the C2C12 cells in DMEM high glucose medium with 20% glucose OR using RPMI to culture ( the cells survive better in this medium) .
2.) for differentiation they let the cells get about 70-80% confluent, for differentiation they replace the serum to 2% horse serum
3.) it is important to use a low passage number (
Make sure the media has glutamine.
I have differentiated C2C12 in DMEM 2% horse serum 4% horse serum and 2% FBS.
the most important aspect is the quality on your cells.
Don't let your cells become to confluent!
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