心肌细胞培养环境对骨髓间充质干细胞定向分化成心肌样细胞有促进作用  

An experimental study on inducing bone marrow mesenchymal stem cell to differentiate into cardiomyogenic cells in cardiomyocyte cultered microenviroment in vitro

作　　者：苏恩本[1] 陈椿[2] 龚觉晓[2] 卢新政[2] 陈子庆[1] 夏新辉[1] 黄峻[2] 马文珠[2] 

机构地区：[1]南京医科大学第一附属医院中心实验室,江苏南京210029 [2]南京医科大学第一附属医院心脏科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2007年第12期1387-1389,1470,I0002,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30370575/C03030201);江苏省教育厅自然科学基金资助项目(04KJB10086)

摘　　要：目的:研究心肌细胞培养微环境对骨髓间充质干细胞(MSCs)定向分化成心肌样细胞的影响。方法:取SD大鼠骨髓,通过贴壁法进行骨髓间充质干细胞培养,经多次传代后获得较纯的MSCs。用5-氮胞苷(5-aza)进行诱导分化,实验组为在玻片上的5-aza干预的MSCs移入到心肌细胞培养环境中;对照组为5-aza干预的MSCs。显微镜下观察两组形态变化,并通过免疫组化鉴别心肌心肌特异蛋白Desmin和肌钙蛋白I(cTnI)表达及其出现时间的早晚。结果:对照组MSCs诱导后10天可见细胞由原来扁平多角形变成长梭形,15天后部分细胞体明显增粗,可见到有些细胞间有融合样情况发生,20天左右类似肌管样细胞出现。而实验组在17天左右可以看到类似心肌管样细胞。免疫组化表明实验组的阳性结果出现的时间比对照组要早。结论:心肌细胞培养环境对骨髓间充质干细胞定向分化成心肌样细胞有促进作用。Objective：To study the effect of cardiomyocyte microenviroment in vitro on bone marrow mesenchymal stem cell（MSCs） in diffierentiating into cardiomyogenic cell. Methods：Bone marrow cells were isolated form SD rat＇s leg bone by their adherence to plastic in vitro, the MSCs were purified by culture-expanded and passaged. After treaded with DNA demethylating agent 5-azacytidine （5-aza）. In experiment group the MCSs were grown,which were cultured in dishes seeded with cardiomyocytes from neonatal rats. In control group the MSCs were cultured only. The MSCs differentiation was observed by microscope,immunohistochemistry staining for Demin and cardiac troponin Ⅰ （cTnⅠ）,and the positive time were observed. Results：In control group,alter treated with 5-aza for 10 days, the clones of adhenrent fibroblast-like cells were formed,and they connected with adjoining cells 15 days and formed myotube- like structures about 20 days later. But the experiment group they formed myotube-like structures alter about 17 days. The results of immunohistochemistry staining indiated that the experiment group＇s positive time was earlier than the control group＇ s. Conclusion： The cardiomyocyte microenviroment are thought to be a key factor in inducing bone marrow mesenchymal stem cells to differentiate into cardiomyogenic cells.

关 键 词：心肌细胞微环境 骨髓间充质干细胞 5-氮胞苷 心肌样细胞 

分 类 号：R329.3[医药卫生—人体解剖和组织胚胎学]