CD105^＋滑膜间充质干细胞成软骨能力

Chondrogenic capacity of CD105-positive synovium-derived mesenchymal stem cells

目的 分选CD105^＋滑膜间充质干细胞（SMSCs）,观察其增殖和向软骨细胞分化的能力.方法 酶消化滑膜组织分离SMSCs,流式细胞仪分选CD105^＋ SMSCs;第3、7天采用WST-1测定SMSCs的增殖能力;软骨诱导21 d进行免疫组织化学染色,检测蛋白聚糖和Ⅱ型胶原.结果 酶消化获取的SMSCs为星形或梭形,分选后SMSCs形态无明显变化.免疫荧光显示分选组CD105^＋细胞较未分选组明显增多.WST-1增殖检测提示两组细胞吸光度值3 d（0.376±0.012、0.329±0.012）、7 d（0.581±0.009、0.524±0.007）比较差异有统计学意义（P＜0.05）.成软骨诱导培养21 d后,分选组甲苯胺蓝和Ⅱ型胶原染色均较未分选组多且深,说明CD105^＋ SMSCs合成更多软骨细胞外基质.结论 CD105^＋ SMSCs具有较强的增殖和成软骨能力,CD105^＋ SMSCs可成为软骨组织工程良好的种子细胞.

Objective To evaluate the proliferation and chondrogenic differentiation of CD105^＋ enriched synovium-derived mesenchymal stem cells （SMSCs）.Methods The synovium-derived mesenchymal stem cells （SMSCs） were obtained from human synovium by enzyme digestion and CD105^-positive （CD105^＋） ceils were enriched using fluorescence activated cell sorting （FACS）.The proliferation ability of CD105^＋ SMSCs was measured through WST-1 at day 3 and day 7.The chondrogenic differentiation of CD105^＋ SMSCs was detected by toluidine blue staining for aggrecan and immunohistochemistry staining for type Ⅱ collagen after 21 days introduction.Results The synovium-derived mesenchymal stem cells （SMSCs） showed star or spindle shape.No obvious differences in morphology were noted after sorting.Immunofluorescence staining showed that there were more CD105^＋ cells in the sorted group than the unsorted.There was statistical difference between sorted and unsorted cells at day 3 （0.376 ± 0.012,0.329±0.012） andday7 （0.581 ±0.009,0.524 ±0.007） on theA value by WST-1 （P〈0.05）.The toluidine blue staining for aggrecan and immunohistochemistry staining for type Ⅱ collagen were more and stronger in sorted cells than unsorted after 21 days chondrogenic differentiation,which indicated that CD105^＋ SMSCs synthesized more cartilage extracellular matrix.Conclusion The CD105^＋ SMSCs have stronger proliferation ability and chondrogenic capacity,which may be promising cell sources for cartilage tissue engineering.