人髌下脂肪垫干细胞与骨关节炎软骨细胞间接共培养可促进其向软骨细胞分化

发布时间：2018-11-10 20:42

【摘要】：目的将人髌下脂肪垫干细胞(IPFPSC)及骨关节炎软骨细胞(OAC)进行间接共培养,促进OAC软骨表型的恢复及诱导IPFPSC向软骨细胞分化。方法体外分离、培养人IPFPSC及OAC,分为单独IPFPSC组、单独OAC组及IPFPSC和OAC间接共培养组,体外成软骨诱导21 d后,通过HE染色、Alcian蓝染色、免疫荧光细胞化学染色检测细胞1型胶原蛋白(Col1)、Col2、Col10、聚集蛋白聚糖(aggrecan)及SOX-9的表达情况。结果共培养组OAC聚集成微球,IPFPSC呈椭圆形;单独组OAC聚集分布,排列松散,单独组IPFPSC呈长梭形。HE染色显示共培养组OAC聚集成团,核染色深,IPFPSC呈椭圆形,胞质丰富,呈旋涡状生长;单独组OAC聚集分布,核染色浅;单独组IPFPSC呈长梭形,核染色浅。Alcian蓝染色显示共培养组OAC及IPFPSC蛋白多糖分泌多,单独OAC及单独IPFPSC组分泌少。免疫荧光细胞化学染色显示共培养组IPFPSC及OAC中Col2、aggrecan及SOX-9表达较强,Col1与Col10表达较弱;而单独培养的IPFPSC及OAC组则相反。结论 IPFPSC和OAC间接共培养可以促进OAC软骨细胞表型的恢复及诱导IPFPSC向软骨细胞分化。
[Abstract]:Objective to promote the recovery of OAC cartilage phenotype and induce the differentiation of IPFPSC into chondrocytes by indirect co-culture of human infrapatellar fat pad stem cells (IPFPSC) and osteoarthritis chondrocytes (OAC). Methods Human IPFPSC and OAC, were isolated and cultured in vitro. Human IPFPSC and OAC, were divided into IPFPSC group, OAC group and IPFPSC and OAC coculture group. After 21 days of chondrogenesis in vitro, HE staining and Alcian blue staining were used. The expression of type 1 collagen (Col1), Col2,Col10, aggregation proteoglycan (aggrecan) and SOX-9 were detected by immunofluorescence cytochemical staining. Results in co-culture group, OAC gathered into microspheres and IPFPSC was oval. The aggregation distribution of OAC in single group was loose and the IPFPSC in single group was long fusiform. HE staining showed that OAC in coculture group was clustered into a cluster with deep nuclear staining, elliptical IPFPSC, rich cytoplasm and swirling growth, and OAC aggregation distribution in single group was shallow in nuclear staining. Alcian blue staining showed that OAC and IPFPSC produced more proteoglycan in coculture group than in OAC and IPFPSC group. Immunofluorescence cytochemical staining showed that the expression of Col2,aggrecan and SOX-9 in IPFPSC and OAC was stronger than that of Col1 and Col10 in co-cultured group, but the expression of Col1 and Col10 in IPFPSC and OAC group was opposite. Conclusion indirect co-culture of IPFPSC and OAC can promote the phenotype recovery of OAC chondrocytes and induce IPFPSC to differentiate into chondrocytes.
【作者单位】：湖南师范大学第一附属医院;第三军医大学西南医院关节外科;
【基金】：国家自然科学基金重点项目(31130021);国家自然科学基金青年项目(31300773)
【分类号】：R684.3

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