滑膜间充质干细胞与软骨细胞接触式共培养构建软骨的初步研究  

Direct Co-culture of Synovial Stem Cells and Chondrocytes Induces Enhanced Cartilage Tissue Formation

作　　者：邵博[1] 胡露露[1] 龚忠诚[1] 杨萌[1] 宁晓婷[1] 王玥[1] 胡鑫[1] 刘慧[1] 克热木.阿巴斯 凌彬[1] 尹小朋[1] 高芸竹 林兆全[1] 

机构地区：[1]新疆医科大学第一附属医院颌面肿瘤外科新疆医科大学口腔医学院新疆维吾尔自治区口腔医学研究所,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院中心手术室,新疆乌鲁木齐830054

出　　处：《口腔医学研究》2016年第6期554-558,共5页Journal of Oral Science Research

基　　金：国家自然科学基金项目(编号:31260229);新疆维吾尔自治区优秀青年科技创新人才培养项目(编号:2014721046)

摘　　要：目的:探讨大鼠软骨细胞与滑膜间充质干细胞以1∶1比例接触式共培养,评价软骨细胞诱导干细胞在体外成软骨方向分化。方法:相同培养基体外培养大鼠滑膜间充质干细胞与软骨细胞,扩增至第3代滑膜间充质干细胞与第1代软骨细胞按1∶1比例混匀,以1×106/mL为终浓度的微团体外培养作为实验组,以相同终浓度细胞数的软骨细胞和滑膜间充质干细胞作为对照组,每组各接种6组。各组标本于体外培养21d后,通过形态学观察,组织学染色,RT-PCR检测产物等方法对其新生软骨进行评价。结果:实验组及对照组体外培养21d后,形成微团似软骨样组织,质地较韧,乳白色。免疫组化法进行Ⅱ型胶原鉴定,基质能被Ⅱ型胶原染色。RT-PCR结果显示诱导后的微团表达软骨特异性基因Ⅱ型胶原和蛋白聚糖。结论:滑膜间充质干细胞与软骨细胞通过接触式共培养形成较成熟的软骨。Objective：To discuss direct co-culture of rat chondrocytes and synovial-derived mesenchymal stem cells at 1：1and evaluate chondrocytes induced differentiation of stem cells in vitro.Methods：Rat synovial-derived mesenchymal stem cells and chondrocytes were cultivated in vitro in the same medium.The 3rd generation of synovial-derived mesenchymal stem cells and the 1st generation of chondrocytes in the proportion of 1：1were mixed.The pellet at the final concentration of 1×106/mL was set as the experimental group,and the considerable population of chondrocytes and synovial-derived mesenchymal stem cells was set as the control group.After cultivated in vitro for 21 days,the newly-born cartilage was evaluated through morphological observation,histological stain,and RT-PCR detection products.Results：After cultivated for 21 days,pellet chondroid tissue was formed and presented tenacious texture and milk white.Type-II collagen was identified by immunohistochemical staining.Matrix could be stained by Type-II collagen.RT-PCR results indicated that pellet expressed cartilage specific genes Type-II collagen and proteoglycan.Conclusion：Through direct co-culture,synovial-derived mesenchymal stem cells and chondrocytes can form mature cartilage.

关 键 词：滑膜间充质干细胞 软骨细胞 共培养 组织工程 

分 类 号：R684[医药卫生—骨科学]