两种方法分离培养类风湿性关节炎成纤维样滑膜细胞比较.docVIP

两种方法分离培养类风湿性关节炎成纤维样滑膜细胞比较[摘要] 目的 比较体外分离培养成纤维样滑膜细胞（FLS）的方法。 方法 滑膜组织来自关节镜清洗术的活动期类风湿关节炎患者，分别采用消化酶培养法、组织块培养法分离培养FLS。采用倒置相差显微镜以及流式细胞术对第3～4代FLS进行鉴定。 结果 2种原代分离培养的方法均可成功培养出FLS。传代可使FLS达到形态学上的纯化要求，倒置相差显微镜观察均符合FLS的形态结构特征，流式细胞术检测示CD44以及CD90呈强阳性，CD55呈弱阳性。组织块法培养法FLS成功率较高，细胞游出时间较早，细胞损伤小。 结论 组织块培养法适合于FLS的培养，第3～4代细胞的数目、活性及纯度符合进一步实验的要求。 [关键词] 成纤维样滑膜细胞；细胞培养；分离培养；类风湿性关节炎 [中图分类号] R-33 [文献标识码] A [文章编号] 1674-4721（2012）07（c）-0013-03 Comparative of two methods for rheumatoid arthritis fibroblast-like synovial cells isolation CAI Wenhong1 SUN Baodong1 ZHANG Baofeng2 CHENG Wenxiang3 YUE Ye3 HU Yiping3 LI Jinchao3 ZHANG Peng3 1.Department of Rheumatology, Shenzhen People′s Hospital in Guangdong Province, Shenzhen 518020, China; 2.Department of Nutrition, Shenzhen People′s Hospital in Guangdong Province, Shenzhen 518020, China; 3.Center for Translational Medicine RD, Shenzhen Institute of Advanced Technology, Chinese Academy of Science, Guangdong Province, Shenzhen 518055, China [Abstract] Objective To compare the isolated methods for fibroblast-like synoviocytes (FLS). Methods Synovium tissue obtained from patients with active rheumatoid arthritis by digestive enzyme culture and tissue culture. The 3～4 generation FLS were identified by morphology and flow cytometry. Results These two methods could culture FLS successfully, tissue culture method was better than the digestive enzyme culture method. FLS showed typical morphological characters under inverted phase contrast microscope. CD44 and CD90 were strong positive, and CD55 was weak positive in the FLS cell surface. Tissue culture method had higher successful rate with earlier and more quickly cell emigration from synovium tissue. Conclusion Tissue culture method is especially suitable for FLS culture in vitro from synovium tissue. And purity of FLS of the third or forth generation meet the requirement for molecular biology experiments. [Key words] Fibroblast-like synoviocyte; Cell culture; Separate training; Rheumatoid arthriti