大鼠心肌细胞诱导人骨髓间充质干细胞向心肌样细胞分化的研究  

The study about the cardiac differentiation of human mesenchymal stem cells induced by rat cardiomyocytes

作　　者：常静[1] 雷寒[1] 陈建斌[2] 黄仁英 

机构地区：[1]重庆医科大学附属第一医院心内科,重庆400016 [2]重庆医科大学附属第一医院血液内科,重庆400016 [3]垫江县妇幼保健院,重庆648300

出　　处：《重庆医科大学学报》2007年第9期907-910,共4页Journal of Chongqing Medical University

基　　金：WHO奖学金资助项目(2004)

摘　　要：目的:研究人间充质干细胞(MSCs)向心肌方向分化的机制。方法:将人MSCs以10000/㎝2的密度接种,共分为四组:1)对照组;2)共同培养组:人MSCs和新生SD大鼠心肌细胞进行共同培养;3)条件培养液组:将从新生SD大鼠心肌细胞培养中获得的培养液,用于人MSCs的培养;4)细胞匀浆液组:将心肌细胞匀浆液用于人MSCs的培养。分别于实验第2、5、7天收集各实验组的人MSCs,进行Desmin及肌钙蛋白I的免疫组化染色。在共同培养组中,同时还用抗人DNA进行的荧光原位杂交,以区分鼠来源的细胞及人来源的细胞。结果:在对照组、条件培养液组及细胞匀浆液组均未检测到心肌特异性蛋白的表达。在共培养组中,第2天,部分人MSCs开始表达Desmin,第5天细胞开始表达cTnI,至第7天阳性表达率达35%左右。结论:与大鼠心肌细胞共同培养可以诱导人MSCs向心肌方向的分化,而心肌细胞的条件培养液及心肌细胞匀浆不能诱导它们的心肌分化。Objective： To explore the mechanism about the cardiac differentiation of human mesenchymal stem cells（MSCs） induced by rat cardiomyocytes. Methods：To induce the cardiac differentiation of human MSCs by using rat cardiomyocytes in different ways （three dishes for each way）：（1） Cocultured with rat cardiomyocytes at the ratio of l：l;（2）Cuhured with the conditional medium,which got from the culture of rat cardiomyocytes and mixed with fresh medium by 1：1;（3） Cultured with the supernatant,which got from the minced rat cardiomyocytes and mixed with fresh medium by 1： 10. The human MSCs were harvested at the day of 2,5 and 7, respectively.Then the cells were stained with desmin and cardiac troponin I monoclonal-antibodies,and fluorescent in situ hybridization to human DNA was used to mark the cells from human. Results： No desmin or cTnI was detected in the human MSCs cultured with conditional medium or supernatant. While for the human MSCs cocultured with rat cardiomyctes,desmin was detectable at the day of 2 and cTnI at the day of 5. Until the day of 7, about 35% of cocultured-human MSCs expressed those cardiac specific antigens. Conclusion：Human MSCs could be induced to differentiate into cardiomyocyte-like cells by coculture with rat cardiomyocytes,but not by the conditional medium or the supernatant of rat cardiomyocytes.

关 键 词：间充质干细胞 心肌细胞 共同培养 心肌分化 

分 类 号：R541.4[医药卫生—心血管疾病] R318.11[医药卫生—内科学]