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updated with more media details
==Growth and Maintenance==
*IMPORTANT - DO NOT ALLOW CULTURES TO BECOME CONFLUENT. Cultures must not be allowed to become confluent as this will deplete the myoblastic population in the culture.
*Split cells normally 1/10 or 1/20 into 10% FBS in DMEM with 1x PSG (see [[ Splitting Cells ]])*Try to split at 70-80% confluence. Healthy growing cells will reach confluence every other day after a 2X dilution. Split them as fibroblasts into the final format (12 well plate, 6 well plate etc.)*Cells normally need to be split every other dayand we maintain them in 100 mm dishes, if they are not ready to split, refeed them on the second day.
==Differentiation==
*When cells reach 80-90% confluence switch to media (DMEM, 1x PSG with 2% horse serum).*Replenish with fresh horse serum media media every other day.*Myotube formation is stimulated when the medium is supplemented with 2% horse serum instead of fetal bovine serumCells should be fully differentiated into myotubes by day 7.
see http://www.stanford.edu/group/blau/protocols/c2lineprotocol.html and http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=crl-1772&Template=cellBiology for more details.