人滑膜间充质干细胞的分离培养与鉴定  

Isolation and identification of human synovial mesenchymal stem cells

作　　者：陈松[1] 符培亮[1] 丛锐军[1] 吴海山[1] 许震宇[2] 

机构地区：[1]上海长征医院关节外科,上海200003 [2]第二军医大学组织胚胎学教研室,上海200433

出　　处：《中国骨与关节外科》2014年第2期146-151,共6页Chinese Journal of Bone and Joint Surgery

基　　金：国家自然科学基金青年科学基金资助项目(81000798)

摘　　要：背景:软骨一旦损伤将很难自我修复,组织工程学修复损伤软骨是目前的研究热点。组织工程学中找到合适的"种子细胞"至关重要。滑膜间充质干细胞(SMSCs)因较其他来源的间充质干细胞成软骨能力、增殖能力更强,且细胞容易获取、对机体损伤小等优点,越来越受到研究者的青睐。目的:探讨人SMSCs体外分离、培养的方法与步骤,探索对SMSCs进行鉴定的最新方法。方法:关节镜下获取11例行关节镜手术患者的滑膜组织,按照Pei等[6]的方法分离培养滑膜干细胞;从细胞形态学观察、细胞生长曲线分析、CCK-8测定增殖活力、流式细胞仪检测细胞周期和细胞表面抗原、对SMSCs进行成脂/成骨/成软骨诱导、免疫细胞荧光染色以及RT-PCR检测成脂/成骨/成软骨相关基因的表达等方面对SMSCs进行鉴定。结果与结论:按照Pei等的方法可以成功地从人滑膜组织中分离出干细胞,分离出的干细胞具有间充质干细胞特异性表型和多向分化潜能。Background：It is difficult to repair the ininjuried cartilage, and it is a hot spot to repair the injuried cartilage by tissue engi-neering. An appropriate seed cell is crucial in ttissue engineering. Synovial mesenchymal stem cells （SMSCs） is easy to dif-ferentiate into cartilage than other source of mesenchymal stem cells. Meanwhile, SMSCs can be harvested easily. Objective：To study the method and procedure of the isolation and culture of human SMSCs in vitro and investigate the last-est methods of SMSCs identification. Methods： Synovial tissues were harvested from 11 patients undergoing arthroscopic surgery in our hospital. SMSCs were isolated and cultured according to Pei＆#39;s method[6]. The SMSCs were identificated from morphological observation, cell growth curve, proliferation activity determined by CCK-8, flow cytometer, induction of adipo-genic/osteogenicondrogenic immunofluorescence staining and RT-PCR. Results and conclusions：SMSCs were successfully isolated from synovial tissue and had specific phenotype and potential multi-directional differentiation.

关 键 词：滑膜间充质干细胞 分离培养 鉴定 成脂诱导 成骨诱导 成软骨诱导 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]