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作 者:邵博[1] 贺多敏[1] 龚忠诚[1] 王玥[1] 杨萌[1] 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
出 处:《口腔医学研究》2017年第4期353-357,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:对比使用软骨上清液和转化生长因子两种方法诱导滑膜间充质干细胞向软骨细胞分化。方法:分别采用消化法获取SD大鼠滑膜间充质干细胞和软骨细胞。体外扩增。实验组一:通过软骨细胞上清液诱导滑膜间充质干细胞向软骨方向分化。实验组二:通过软骨诱导液(TGF-β1,ITS+Premix,2-磷酸抗坏血酸等)诱导滑膜间充质干细胞向软骨方向分化。培养21d后通过形态学、免疫组织化学法检测其生物学特性,RT-PCR检测诱导后产物Ⅱ型胶原RNA含量。结果:2种诱导方法均能诱导滑膜间充质干细胞成软骨细胞方向分化。在形态学可见2组诱导后产物成软骨样结构,呈乳白色,质地韧。免疫组织化学鉴定基质能被Ⅱ型胶原染色,细胞染色呈现棕黄色。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:两组实验组均能诱导滑膜间充质干细胞向软骨方向分化。对比两种实验方法,使用上清液诱导滑膜间充质干细胞向软骨方向分化能力更强。Objective:To study the role of secreted factors by chondrocytes to induce chondrogenesis of synovial mesenchymal stem cells.Methods:The synovial mesenchymal stem cells(SMSCs)and chondrocytes were harvested from rat knee joints and cultured through the digestion method.The supernatant was collected from chondrocytes,centrifuged,filtered,and cryopreserved.Group1:the third passage SMSCs centrifuged as pellets were cultured in the chondrocyte supernatant for 21 days.Group2:SMSCs differentiated under the induction of transforming growth factor beta-1in the same time.The cells morphology was examined and the type II collagen and aggrecan were detected through immunohistochemistry and RT-PCR.Results:The SMSCs induced became cartilage-like tissue after 21 days.The type II collagen was detected positively in the matrix of two groups immunohistochemically.Two groups for chondrogenic differentiation of SMSCs successfully induced the expression of multiple cartilage-specific molecules,including collagen type II and aggrecan,and resulted in a chondrocyte-like phenotype.The Group 1's cartilage gene content was higher than that of the group used transforming growth factor.Conclusion:Two groups can induce SMSCs differentiation into chondrocytes.Using supernatants can induce better results.
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