C2C12 cells when exposed to 2% horse serum are reported to differentiate into myotubes. I have tried 2% horse serum DMEM for 5-6 days but no myotube formation is evident. Can any one suggest the exact composition of differentiating media?
Mayank Verma
Do I need to add ITS to 2%Horse Serum DMEM ? If yes, in what concentration?
Hi.
Where does C2C12 derive from?
If C2C12 once confluent when cultured in myoblast state with 10% FBS, it will never again It will not differentiate to myotubes. I faced the same problem as you, I would recommend making a frozen stock at about 40-60% confluent with ATCC derivatives.Note that some of the commercial C2C12 seems to be mixed once confluent. In that case, give up and try another origin. At least the ATCC ones are sure; the same can also be applied to 3T3L1.
I have tried numerous lots of horse serum and have not found any particular differences between manufacturers and lot. I have never done differentiation enhancement with the addition of Insulin-Transferrin-Selenium because I intended the experiment to include insulin signaling.
Good luck!
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