THE STUDY OF DIRECT DIFFERENTIATION OF MICE EMBRYONIC STEM CELLS INTO CARDIOMYOCYTES INDUCED BY TGF-?_1 AND CO-CULTURE WITH VISCERAL ENDODERM LIKE END-2 CELLS / 解剖学报

ABSTRACT

Objective To study the differentiation of embryonic stem cells(ESCs) induced by transforming growth factor-?_1(TGF-?_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-?_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 ?-sarcmeric actin(?-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-?_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 ?-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