Characterization of myogenic cell lines derived by 5-azacytidine treatment

Three myogenic clonal cell lines were isolated from C3H 10T1/2 C18 cells (10T1/2) treated with 5-azacytidine (5-aza-CR). These lines reproducibly underwent fusion at confluence into functional myotubes capable of contracting in response to acetylcholine. The degree of fusion could be increased two- to threefold if the cells were grown on gelatin-coated dishes. All of the cell lines lost some of their myogenic potential after repeated passaging and the percentage of colonies capable of forming muscle was not increased by permissive media containing 2% horse serum. The 10T1/2 cells expressed only the BB form of creatine phosphokinase but all of the myogenic clones expressed additionally the MM and MB forms of the isozyme after fusion. The overall genomic level of 5-methylcytosine was decreased in some but not all of the cell clones tested. Comparisons between the 10T1/2 cells which never form muscle without 5-aza-CR treatment and clonal derivatives of committed cell types might be of value in understanding the molecular basis of the commitment process.