体外诱导兔骨髓间充质干细胞向心肌细胞分化的研究

目的 :探讨体外培养骨髓间充质干细胞的方法以及定向诱导分化成心肌样细胞的条件。方法 :常规骨髓穿刺获取细胞悬液 ,应用Percoll液密度梯度离心以及贴壁培养技术分离间充质干细胞。使用不同浓度的 5 -氮胞苷定向诱导分化 ,应用透射电镜观察细胞超微结构和免疫细胞化学技术检测细胞特异性心肌收缩蛋白、舒张蛋白以及连接蛋白的表达。结果 :在 5 aza诱导后第 2 0天 ,约 3 0 %的细胞表达肌钙蛋白T、α 横纹肌肌动蛋白、肌浆网磷酸受钙蛋白和连接蛋白 43。透射电镜观察 ,胞浆内粗面内质网丰富 ,部分内质网正在合成肌丝。结论 :骨髓间充质干细胞具有分化为心肌样细胞的潜能。

Bone Marrow Mesenchymal Stem Cells in Rabbit can be Induced Myocardial Cells in Vitro

Objective: To fine out the way of proliferating bone marrow mesenchymal stem cells and investigate the potency of MSCs to differentiation into cardiomyocytes in vitro. Method: MSCs were aspirated from adult rabbits' tibias , isolated by Percoll gradient,then treated with different concentration of 5 azacytidine for 24 hours at the third day of seeding. After 20 days, MSCs differentiation was observed by immunohistochemistry staining and electron microscopy.Results: After co cultured with 10μmol/L 5 azacytidine for 24 hours, myofilaments can be seen by electron microscopy. About 30% cells were found positively stained with cardiac troponin T, α sarcomeric actin,phospholamban and connexin 43 in culture at the 20th day.Conclusion: Bone marrow mesenchymal stem cells have the potency of self expansion and differentiation into cardiomyocytes in vitro.