体外利用慢病毒介导BMP-2基因诱导兔滑膜间充质干细胞向软骨细胞分化  

Lentiviral-medicated BMP-2 gene transfer induces chondrogenesis of rabbit synovium-dervied mesenchymal stem cells in vitro

作　　者：符培亮 张雷 吴海山 吴宇黎 胡凯猛[2] 丛锐军 陈松 

机构地区：[1]上海市长征医院关节外科,200003 [2]第二军医大学组织胚胎学教研室,200433

出　　处：《中华关节外科杂志（电子版）》2013年第4期60-65,共6页Chinese Journal of Joint Surgery(Electronic Edition)

基　　金：国家自然科学基金青年科学基金项目(81000798)

摘　　要：目的使用慢病毒介导兔BMP-2基因转染兔滑膜间充质干细胞(SMSCs),检测其安全性,并在体外定向诱导至软骨细胞。方法通过构建包含BMP-2基因的慢病毒载体,转染SMSCs后行安全性分析;各项检测和MicroRNA分析判断转染后的SMSCs是否进入软骨细胞分化谱系。结果贴块法获得的SMSCs在经分离纯化后,表现出间充质干细胞应有的结构和表面标志物,具有多向分化潜能。增殖动力学、核型分析、致瘤型分析等证实被慢病毒转染的SMSCs安全。Western blot、免疫荧光等证实且能稳定表达BMP-2蛋白。14 d后,免疫组化、MicroRNA检测等证实在不需外加外源性BMP-2的体外环境下SMSCs可自发向软骨细胞分化。结论经重组慢病毒载体感染的兔SMSCs足够安全,能稳定表达BMP-2,体外能自发向软骨细胞分化。Objective To induce chondrogensis differentiation of rabbit synovium-dervied mesenehymal stem ceils （SMSCs） by the recombinant lentiviral medicated BMP-2 gene in vitro, and to test the safety of this method. Methods The lentivirus vector containing BMP-2 gene was constructed and was used to transfect SMSCs. Safety analysis was peformed. Many tests and microRNA analysis were carried out to determine whether infeeted-SMSCs differentiate into chondrocytes. Results SMSCs expressed surface markers of stem cells, and had multi-differentiation potential. The infected SMSCs were safe according to cell proliferation kinetics analysis, karyotype analysis and tumorigenicity analysis. Western blot and immunofluorescence demonstrated SMSCs expressed BMP-2 continually and stably. After culturing the 14 days, SMSCs succeeded in differentiating into cartilage without adding BMP-2 repeatedly, which was verified by immunohistoehemistry and microRNA analysis. Conclusions SMSCs infected by plentiV6- oBMP-2 can express BMP-2 continually and stably, which can promote SMSCs differentiating into chondrogenic lineages spontaneously.

关 键 词：滑膜 间质干细胞 慢病毒属 骨形态发生蛋白质类 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]