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

目的使用慢病毒介导兔BMP-2基因转染兔滑膜间充质于细胞（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。Westernblot、免疫荧光等证实且能稳定表达BMP-2蛋白。14d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。

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

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.