关键词: END-2细胞, 胚胎干细胞, 心肌细胞, 分化, 共培养, BR>免疫荧光技术, 小鼠 BR>

Abstract: Objective To study the differentiation of embryonic stem cells(ESCs) induced by transforming growth factor-β1(TGF-β1) 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-β1 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-β1, 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)%.(P<0.01),and the beating areas were bigger,and more

Key words: END-2 cell, Embryonic stem cells, Cardiomyocytes, Differentiation, Co-culture, Immunofluoresence staining, Mice