人滑膜间充质干细胞的分离培养与鉴定 |
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引用本文: | 陈松,符培亮,丛锐军,等. 人滑膜间充质干细胞的分离培养与鉴定[J]. 中国骨与关节外科, 2014, 0(2): 146-151 |
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作者姓名: | 陈松 符培亮 丛锐军 等 |
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作者单位: | [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 |
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基金项目: | 国家自然科学基金青年科学基金资助项目(81000798) |
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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。
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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 |
Isolation and identification of human synovial mesenchymal stem cells |
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Affiliation: | CHEN Song, FU Peiliang, CONG Ruijun, WU Haishan, XU Zhenyu (1. Department of Joint Center, Shanghai Changzheng Hospital, Shanghai 200003; 2. Section of Histology & Embryology, Basic Research Department, Second Military Medical University, Shanghai 200433, China) |
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Abstract: | 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'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. |
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Keywords: | Synovial mesenchymal stem cells Isolation Identification Adipogenic differentiation Osteogenic differentia-tion Chondrogenic differentiation |
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