Ⅱ型胶原诱导的关节炎大鼠滑膜成纤维细胞的培养、纯化和鉴定

发布时间：2018-03-20 06:12

  本文选题：滑膜成纤维细胞　切入点：胶原诱导的关节炎大鼠模型　出处：《南京师大学报(自然科学版)》2017年03期 　论文类型：期刊论文

【摘要】：类风湿关节炎(Rheumatoid arthritis,RA)病人滑膜成纤维细胞(Fibroblast-like synoviocytes,FLS)是研究RA发病机制和进行RA药物筛选的最理想的细胞来源.但是对于众多非医疗机构的高校和科研院所而言,由于RA病人滑膜组织不易获取而难以进行细胞水平的操作.因此,建立胶原诱导的关节炎大鼠(Collagen-induced arthritis,CIA)模型,成功分离膝关节滑膜组织并进行滑膜成纤维细胞的原代培养和纯化,将有效解决RA FLS的来源问题,为后续实验奠定基础.本实验通过皮内和皮下两次注射鸡Ⅱ型胶原诱导Wistar大鼠罹患关节炎,通过关节炎指数评分和膝关节HE染色明确CIA模型的建立;分离膝关节滑膜组织,采用Ⅱ型胶原酶和无EDTA胰酶消化法分离细胞并进行传代培养,传至第三代时,细胞免疫荧光检测细胞中VCAM-1蛋白含量以鉴定细胞纯度.结果显示,滑膜组织经过消化和培养后得到大量大鼠滑膜成纤维细胞;在显微镜下观察所得细胞呈梭形树突状,具有成纤维样细胞特征;VCAM-1蛋白阳性细胞达98%以上,确证所分离细胞为滑膜成纤维样细胞,且纯度极高.本研究建立了较简便快捷地获得大量滑膜成纤维细胞的方法,为RA的发病机制研究和药物防治提供了丰富的实验材料.
[Abstract]:Fibroblast-like synoviocytes-FLSs (Fibroblast-like synoviocytes-FLSs) from patients with rheumatoid arthritis (Rheumatoid arthritis RAA) are ideal cell sources for studying the pathogenesis of RA and screening RA drugs. But for many non-medical institutions and research institutes, Because synovial tissue of RA patients is difficult to obtain, it is difficult to operate at cell level. Therefore, Collagen-induced arthritis CIAA model of collagen-induced arthritis rat was established, and synovial tissue of knee joint was isolated successfully and primary culture and purification of synovial fibroblasts were carried out. It will effectively solve the problem of the origin of RA FLS and lay a foundation for further experiments. In this experiment, Wistar rats were induced to develop arthritis by intradermal and subcutaneous injection of chicken type 鈪，

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