Soft tissue malignancies often show divergent differentiation, including myogenic lineage. Five rat tumors induced with 20-hydroxymethylcholanthrene (20-OH-MCA), were cultured in vitro, and three cell lines (YMC-1, YMC-2, YMC-3) were established from them. YMC-2 and -3 cells were spindle-shaped, and YMS-1 cells round and epithelioid. In confluency, YMC-3 cells formed myotubes. The nude mouse xenotransplants showed morphological features consistent with their myogenic phenotypes. Muscle-specific enzyme activities were highest in YMC-3 cells. These lines may be useful in the investigation of the myogenic differentiation of undifferentiated mesenchymal cells.