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Primary Myoblast Culture - Could u help me in myoblast culture and differention (Jan/04/2007 )

I am working on primary myoblast culture and differentiation but now I have some problems. One of my big problems is that after thawing the isolated myoblast (from legs of neonatal mice), the morphology and the growth rate of the cells change extremely. So I can not control the cells after thawing. If you have had this kind of experiments, could you give me some advices?
The second question is that is there any one has experiment on myoblast differention into myotube? In my case, I use DMEM (low glucose) + 5%horse serum to differentiate but I can not observe any contractile cell until the day of 20 (the media was changed the other day). And can you tell me the percentage of contractile cells in the dish?

Thank you very much for reading my question. I appreciate all your help and advices.



For myoblast differentiation, I used DMEM high glucose with 0,5% FBS. Some protocols added insuline and apotransferin and various concentration of FBS (from 0,5% to 2%). You can also try to swicht FBS by horse serum. I some cases, it may help to have a better differentiation.