Skeletal muscle cell-derived grafts in the heart may benefit myocardial performance after infarction. Several studies have suggested that skeletal muscle stem cells (satellite cells) from adult muscle undergo transdifferentiation into cardiomyocytes after grafting into the heart, but expression of cardiac markers in graft cells has not been rigorously confirmed. To determine the fate of satellite cell-derived grafts in the heart, adult rat satellite cells were tagged in vitro with bromodeoxyuridine (BrdU) and grafted into normal hearts of syngeneic rats. At 4 and 12 weeks the graft cells formed multinucleated, cross-striated myofibers that expressed fast skeletal myosin heavy chain (MHC), thus indicating a mature skeletal muscle phenotype. Double staining for the BrdU tag and cardiac-specific markers was employed to identify transdifferentiation. Aside from four questionable cells, none of the 11 grafts examined expressed α -MHC, cardiac troponin I, or atrial natriuretic peptide. At 4 weeks, grafts expressed β -MHC, a hallmark of slow twitch myofibers. By 12 weeks, however, the myofibers had atrophied and downregulated β -MHC. Grafts never expressed the intercalated disk proteins N-cadherin or connexin43, hence electromechanical coupling did not occur. In conclusion, satellite cells differentiate into mature skeletal muscle and do not express cardiac-specific genes after grafting into the heart. Thus, transdifferentiation into cardiomyocytes did not occur.