CAJ | 학술논문

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.
목적 분선CD105+활막간충질간세포(SMSCs),관찰기증식화향연골세포분화적능력.방법 매소화활막조직분리SMSCs,류식세포의분선CD105+ SMSCs;제3、7천채용WST-1측정SMSCs적증식능력;연골유도21 d진행면역조직화학염색,검측단백취당화Ⅱ형효원.결과 매소화획취적SMSCs위성형혹사형,분선후SMSCs형태무명현변화.면역형광현시분선조CD105+세포교미분선조명현증다.WST-1증식검측제시량조세포흡광도치3 d(0.376±0.012、0.329±0.012)、7 d(0.581±0.009、0.524±0.007)비교차이유통계학의의(P＜0.05).성연골유도배양21 d후,분선조갑분알람화Ⅱ형효원염색균교미분선조다차심,설명CD105+ SMSCs합성경다연골세포외기질.결론 CD105+ SMSCs구유교강적증식화성연골능력,CD105+ SMSCs가성위연골조직공정량호적충자세포.
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.