滑膜间充质干细胞软骨分化能力的实验研究  

Experimental study of chondrogenesis of synovial mesenchymal stem cells

作　　者：龚忠诚[1,2] 魏丽丽[1] 吴杨[1] 凌彬[2] 刘慧[2] 林兆全[2] 龙星[1] 

机构地区：[1]武汉大学口腔医学院,口腔生物医学工程教育部重点实验室,湖北武汉430079 [2]新疆医科大学第一附属医院口腔颌面外科,新疆乌鲁木齐830011

出　　处：《新疆医科大学学报》2010年第1期22-25,共4页Journal of Xinjiang Medical University

基　　金：新疆医科大学第一附属医院青年基金(2009-QN-8)

摘　　要：目的探讨滑膜间充质干细胞软骨分化能力。方法无菌状态下获取兔颞下颌关节滑膜组织,酶消化法进行滑膜间充质干细胞原代培养,并进行传代扩增。第3代细胞消化后离心成为细胞团块,用软骨诱导液培养,并以鸡尾酒法添加生长因子人重组转化生长因子β1(recombined human transforming growth factorβ1,rhTGF-β1)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF),即前3天2种生长因子共同使用,之后只用rhTGF-β1继续培养18d。细胞团块培养21d后,免疫组织化学染色检测Ⅱ型胶原的表达。细胞团块培养在普通培养基中不添加rhTGF-β1和bFGF作为对照组。结果滑膜间充质干细胞经过传代培养后,成为形态一致的梭形细胞;细胞团块经软骨诱导液、rhTGF-β1和bFGF培养后,免疫组织化学染色可见Ⅱ型胶原在细胞之间的细胞外基质中明显显色,并且细胞团块的周边要比中央浓染;原本二维培养状态下的梭形滑膜间充质干细胞变为球形的软骨样细胞形态。结论滑膜间充质干细胞具有很强的成软骨能力,可以作为髁突软骨组织工程的种子细胞。Objective To explore the differentiation potency of synovial mesenchymal stem cells （SMSCs）. Methods Synovium was harvested from rabbit joints under sterile conditions. SMSCs were obtained and expanded. Passage 3 SMSCs were centrifuged to form pellets. SMSCs pellets were cultured in the chondrogenesis culture medium with cocktail application of recombined human transforming growth factor β1 （rhTGF-β1） and basic fibroblast growth factor （bFGF）. After being cultured 21 days, SMSCs pellets were examined immunohistochemically to evaluate the expression of collagen type Ⅱ. SMSCs pellets cultured in normal culture medium were served as control. Results The phenotype of SMSCs was changed from spindle shape cultured under 2D conditions to round chondrocyte-like shape under 3D conditions. Immunohistochemical collagen type Ⅱwas stained stronger at the outside of SMSCs pellets than at the center. Rich chondroginous extracellular matrix was filled in the inter-cell ular space. Conclusion It is suggested that SMSCs could be served as seeding cells for mandibular condyle cartilage tissue engineering.

关 键 词：滑膜细胞 软骨分化 人重组转化生长因子β1 碱性成纤维细胞生长因子 

分 类 号：R681.3[医药卫生—骨科学] R686.7[医药卫生—外科学]