5-氮胞苷诱导骨髓间充质干细胞向心肌样细胞分化观察

Experimental investigation of 5-azacytidine inducing rat bone marrow mesenchymal stem cells to differentiate into cardiomyocytes

目的:探讨5氮胞苷(5aza)对纯化培养的骨髓间充质干细胞(MSCs)向心肌样细胞分化的影响。方法:取雄性Wistar大鼠下肢长骨骨髓进行体外培养和连续传代,取第3代MSCs以10μmol/L5aza诱导作用24h后,用含体积分数为5%胎牛血清的完全培养基继续培养,观察细胞形态学变化,并在4周后行免疫细胞化学染色鉴定αActin,Desmin及心肌特异性肌钙蛋白T(cTnT)的表达。结果:5aza作用MSCs7d后细胞形态及排列发生变化,14～21d左右细胞数量较诱导前明显减少,28d后细胞稀落,周围伸出多个长的突起,细胞体积大小不等。经抗αActin抗体、抗Desmin抗体、抗cTnT抗体鉴定部分细胞出现阳性染色,阳性细胞率分别为20%,19%和10%。结论:骨髓间充质干细胞在体外特定的诱导条件下可以向心肌样细胞分化,有望成为缺血性心脏病细胞移植治疗的细胞材料。

Aim： To investigate the effects of 5-azacytidine on inducing rat bone marrow mesenchymal stem cells to differentiate into cardiomyocytes. Methods ： The bone marrow of male Wistar rats was separated and purified. The MSCs at passage 3 were induced by 10 μmol/L 5-azacytidine medium. The morphological changes of cells were observed then immunocytoehemistry was used to identify the α-Actin, Desmin and cTnT. ReSults： Cell phenotype and array were changed after being treated by 5-azaeytidine. The cell proliferation was inactive. There were partial cells showed positive stain after 4 week induction by 5-azacytidine. Conclusion ： The MSCs could be induced in vitro to differentiate into cardiomyocytes by 5-azacytidine, which suggests that MSCs might be a novel cell source for cellular transplantation in treatment of ischemic heart disease.