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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 compare the similarities and differences between the four methods in the culture of rheumatoid arthritis (RA) synovial cells (FLS) and to provide a better in vitro experimental model for the study of RA. Methods Twenty cases of synovial tissue samples were isolated and cultivated with four methods of tissue block method, single collagenase digestion, single trypsin digestion and double enzyme digestion. RA FLS were cultured on the 7th day, 4 generation of cell identification, immunohistochemical identification of cells, the number of isolated cells and purity. Results Compared with the culture time of the four culture methods, single collagenase method> double enzyme digestion method> tissue block method> single pancreatic enzyme method (P <0.05). Tissue block method and double enzyme digestion method cells than the single collagenase method and double enzyme digestion less (P <0.05). The fourth generation of cells are mainly FLS (> 95%). Conclusions The four methods can be used to obtain RA FLS cell line with stable biological characteristics. Tissue block method is a rapid method to establish experimental model of RA in vitro with high success rate.