类风湿性关节炎细胞水平建模四种方法比较  

Comparison of 4 methods for isolation and characterization of rheum atoid arthritis synovial fibroblasts from primary culture

作　　者：胡赟霞[1] 齐文成[2] 徐新女[3] 韩锋[2] 

机构地区：[1]天津医科大学,天津300007 [2]天津市第一中心医院 [3]卫生部急救医学重点实验室

出　　处：《山东医药》2009年第16期21-23,共3页Shandong Medical Journal

基　　金：天津市卫生局计划面上项目基金资助项目(05KY11)

摘　　要：目的比较四种方法在培养类风湿性关节炎(RA)滑膜细胞(FLS)中的异同,为研究RA提供较好的体外实验模型建模方法。方法20例滑膜组织标本均用组织块法、单胶原酶消化法、单胰酶温消化法和双酶消化法四种方法分离、培养RA FLS,均取培养第7日细胞计数,取第4代细胞鉴定,免疫组化鉴定细胞,测定分离细胞的数量和纯度。结果四种培养方法的培养时间比较,单胶原酶法>双酶消化法>组织块法>单胰酶法(P<0.05)。组织块法及双酶消化法所得细胞数较单胶原酶法及双酶消化法少(P<0.05)。第4代细胞均以FLS为主(>95%)。结论四种方法体外培养可获得生物学特性稳定的RA FLS细胞系,其中组织块法是细胞水平建立RA体外实验模型成功率较高的方法,单纯胰酶法是较快速的方法。Objective To find an effective and fast method for isolation and characterization of rheumatoid arthritis （RA） fibroblast-like synovioeytes （FLS） in vitro in order to research RA better. Methods Tissue block method, collagenase method, trypsin method and double enzyme method were compared. Calculate main operational time for each method, account number of cells after 7 days cultured with each method, and compare results of immunohistochemistry. Results The sequence of four methods＇ efficacy was tissue block, double enzyme, collagenase, trypsin method in sequence. In time consuming, trypsin was significantly faster than other methods; cell numbers was accounted at the 7th day of primary culture. The immunohistochemistry demonstrated that there was no significantly difference between each methods and cells were most FLS（ 〉95% ）. Conclusion Tissue block method is more effective than others while trypsin method is faster than others.

关 键 词：类风湿性关节炎 滑膜细胞 细胞 培养的 组织块法 单纯胰酶法 

分 类 号：R593.22[医药卫生—内科学] R329.2[医药卫生—临床医学]