The study of direct differentiation of mice embryonic stem cells into cardiomyocytes induced by TGF--beta_(1) and co--culture with visceral endoderm like END--2 cells

Objective To study the differentiation of embryonic stem cells(ESCs) induced by transforming growth factor-β_1(TGF-β_1) and co_cultured with visceral endoderm like END_2 cells,and explore cardiomyocytes induction effects of the combined techniques Methods Day 2-3 embryoid bodies(EBs) were derived from ESCs,and then TGF-β_1 was added or/and co_cultured with END_2 cells or END_2 cells conditioned medium.Spontaneous differentiation was as a control.The expression of cardiac specific α-sarcmeric actin(α-actin) and cardiac troponin_T(TnT) was detected by immunofluoresence staining.The ultrastructural analysis for ESCs_derived cardiomyocytes was scanned by transmission electron micrograph. Results The total percentage of beating EBs treated with TGF-β_1,co_cultured with END_2 cells,or END_2 cell conditioned medium was(43±2.08)%,(69±3.61)%,(65±3.06)%,respectively.All the beating cardiomyocytes derived from ESCs expressed cardiac_specific proteins for α-actin and TnT,and could be observed the cardiac_specific ultrastructure.Interestingly,the total percentage of beating EBs treated with the combined method was(91±1.52)%.(P0.01),and the beating areas were bigger,and more purified cells could be gained.Conclusion By using combination induction method it could achieve a synergistic effect on the differentiation of ESCs into the cardiomyocytes.