人骨髓间充质干细胞体外定向分化为心肌细胞的实验研究

Experimental study on the induced differentiation of human bone marrow mesenchymal stem cells into cardiomyocytes in vitro

目的:建立人骨髓间充质干细胞(MSCs)体外定向分化为心肌细胞的实验模型。方法:取6个月水囊引产胎儿股骨骨髓,Percoll分离液分离,取单个核细胞层在人MSCs专用MSCGM培养基中体外培养。将第3代生长状态良好的MSCs用10μmol/L5-氮胞苷孵育诱导24h,继续向心肌细胞诱导分化。结果:用Percoll分离液(1.073g/ml)分离得到的单个核细胞48￣72h呈纺锤形、梭形、多角形的多型性改变;经5-氮胞苷孵育24h,细胞逐渐向梭形心肌细胞样形态转变,胞浆中可见棕黄色颗粒。结论:MSCs在体外经5-氮胞苷孵育24h,可被定向诱导分化为心肌形态的细胞。

Objective： To establish a model of human mesenchymal stem cells （MSCs） differentiat ed into cardiomyocytes in vitro. Methods： Bone marrow samples were derived from bone marrow of a 6-month fetus aborted with water bag. Mononuclear cells were separated and isolated by gradient centrifugation in 1.073g/ml Percoll solution. Then they were seeded and expanded in specific human MSCs medium MSCs in third passage were co-cultured and treated with 10 μmol/L 5-aza for 24 hours. Results： Mononuclear cells adhered to the flasks and grew typically in fibroblastic or spindle shape. Multifoot cells changed into orange-like ones with a spindle shape as that of cardiomyocytes. By immmunocytochemistry, they were found to be positively stained with actin. Conclusion： Human MSCs can be induced to differentiate into cardiomvocvtes by 5-azacvtidine in vitro.