Study on Inducing Differentiation of Mouse Embryonic Stem Cells into Cardiomyocytes in Vitro

ZENG Bin, LIN Guosheng, LUO Hao, CAI Jun Dept. of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China  

Objective: To analyze the differentiation features of mouse embryonic stem cells(ESCs) induced by TGF-β_ 1 and co-culture of visceral endoderm like (END-2) cell. Methods: Mouse embryonic fibroblast(MEF)feeders were used to promote the growth of ESCs and were maintained in an undifferentiated state. Embryoid bodys EBs aged 2-3 days were derived from ESCs,then were added to 24 holes panel containing TGF-β_ 1 and added to 24 holes panel covered a feeder layer of END-2 cells,containg TGF-β_ 1.The expression of cardiac specific α-sarcmeric actin( α-actin) and cardiac troponin-T(TnT) was detected by using immunofluoresence.The ultrastructural analysis of ESCs-derived cardiomyocytes was scanned by transmission electron micrograph.The percentage of EBs cluster-derived cardiomyocytes was evaluated by flow cytometry. Results: The total percentage of beating EBs treated with TGF-β_ 1 and TGF-β_ 1 combined with END-2 feeder layer was 43% and 91%(P0.05),respectively. All the beating cardiomyocytes derived from ESCs expressed cardiac-specific proteins for α-actin and TnT, and the cardiac-specific ultrastructure could be observed. Interestingly, under the condition of co-culture with END-2 cell,the beating areas were bigger,and more purified cells could be gained. The percentage of EBs cluster-derived cardiomyocytes between the two induction groups was 35% and 74%(P0.05), respectively. Conclusion: Added TGF-β_ 1 co-cultured with visceral endoderm like END-2 cell could achieve a synergistic effect on the differentiation of ESCs into the cardiomyocytes, and could get a higher induction efficiency.

【Key Words】： END-2 cell Embryonic Stem Cell Cardiomyocyte Co-culture Differentiation
【Fund】： 湖北省卫生厅重点项目(编号:HB05336)
【CateGory Index】： Q813.11

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