Differentiation of mouse embryonic stem cells into cardiomyogenic cells in cardiomyocytes

OBJECTIVE To explore the effect of embryonic stem cells (ESCs) differentiating into cardiomyocytes (CM) in cardiomyocytes. METHODS The blastocytes of 3.5d were collected from Kunming species mouse, cultured on mouse embryonic fibroblaste cell feeder layers for 4-5d, and the ESCs were isolated and subsequently cultured in vitro. At the day of 2 to 3, the embryoid bodies (EBs) were derived from ESCs of passage 3 to 5, and then cocultured with neonatal cardiomyocyte (CM) to induce into cardiomyocytes. Then the expression of cardiac specific cardiac troponin-T (TnT) and а-Actin were detected by immunofluoresence. RESULTS The rhythmic beating embryoid bodies were observed on day 3, and reached to 93% of rhythmic beating embryoid bodies on the 12th day in cocultured group. All the beating cardiomyocytes derived from ESCs expressed cardiac-specific proteins for TnT and а-Actin, and the group induced by direct cell to cell contact acquired the highest differentiating ratio of 56.5%, and was the highest differentiating ratio compared with other groups (P﹤0.05). CONCLUSION Car- diomyocyte coculturing system and direct cell-to-cell interaction can induce mouse ESCs to differentiate into cardiomyocyte-like cells.