体外定向化学诱导成人脂肪源性间充质干细胞分化为心肌样细胞

Chemical inducing differentiation of adult adipose-derived mesenchymal stem cells into cardiomyocyte-like cells in vitro

目的:研究在体外不同浓度、不同作用时间诱导条件下5-氮胞苷(5-aza)对脂肪间充质干细胞(ADMSCs)诱导分化为心肌细胞的影响。方法:酶消化筛选法体外分离ADMSCs并传代培养,流式细胞仪鉴定细胞CD44、CD34的表达,不同浓度5-aza(1、5、10、15、20μmol/L)分别诱导12、24、48、72 h,相差显微镜下观察细胞形态变化;诱导后4周时免疫细胞化学鉴定心肌特异性肌钙蛋白I(cTn-I)的表达,分析细胞转化率。结果:体外分离培养的ADMSCs表达CD44,不表达CD34;5-aza诱导后4周时免疫细胞化学显示cTn-I表达阳性,以10μmol/L 5-aza诱导24 h为适宜诱导条件;结论:体外5-aza化学诱导下成人脂肪组织间充质干细胞具有向心肌细胞分化的潜能。

Objective： To study the differentiation of adult adipose - derived mesenchymal stem cells （ADMSCs） into cardiomyoctes in vitro induced by 5 - azacytidine of different concentration for different time length. Metheds： ADMSCs were isolated and cultured from adult adipose tissue, and the expression of CD44 and CD34 were detected by flow cytometry. The cells were induced with medium containing 1, 5, 10, 15, 20 μmol/L 5 - azacytidine for 12, 24, 48, 72 hours respectively, and the morphological changes of those cells were observed by phase -contrast microscope. The immunocytochemistry method was used to detect expression of cardiac troponin - Ⅰto analyze transformation rate of cardiomyocyte - like cells. Results： Flow cytometry showed that the ADMSCs expressed CD44 but not CD34, and the morphology of induced cells was like cardiomyocytes. The induced cells expressed cardiac troponin - Ⅰ 4 weeks after induction by 5 - azacytidine, which demonstrated that the best condition for inducing differentiation of ADMSCs cells into cardiomyocytes in vitro was 5 - azacytidine of 10 μmol/L for 24 hours. Conclusion： This study showed that adult adipose -derived mesenchymal stem cells could be induced to differentiate into cardiomyocyte -like cells by 5 -azacytidine in vitro.