【摘要】 目的探讨人胃癌肿瘤相关成纤维细胞(carcinoma-associated fibroblasts,CAFs)的培养和鉴定方法,并分析其与胃正常成纤维细胞(normal fibroblasts,NFs)在生物学特性和蛋白表达方面的差异,为其功能研究奠定基础。方法采用组织块法和酶消化法进行人胃癌CAFs和NFs的原代培养,通过酶消化法进行人工纯化,通过形态学观察和多种蛋白的免疫细胞化学染色对其进行鉴定。结果获得纯化的人胃癌CAFs和NFs。胃癌CAFs呈长梭形,大小不等,生长密集,排列紊乱。除表达vimen-tin这一成纤维细胞的标记物外,还表达FAP、SMA,但不表达CK、desmin。结论只要给细胞创造良好的生长条件并规范操作,组织块法和消化法均可成功的培养出人胃癌CAFs和人胃NFs。胃癌CAFs与NFs在形态结构、生长方式和蛋白表达等方面均存在明显差异。更多还原

【Abstract】 Purpose To investigate the purification and identification of gastric carcinoma-associated fibroblasts(CAFs).Methods The primary gastric CAFs and normal fibroblasts(NFs) were obtained by tissue culture and digestion methods.Then cells were dissociated and purified by EDTA and trypsinization.The expression of vimentin,SMA,FAP,cytokeratin in the cell was examined by immunocytochemistry.Results The purified gastric CAFs and NFs were maintained.The characteristics of shape and growth of gastric CAFs significantly changed as compared with NFs.The gastric CAFs showed negative staining for cytokeratin and desmin,and positive staining for vimentin,SMA and FAP.Conclusions The primary culture of the fibroblasts in human gastric carcinoma tissue can be performed successfully with digestion method or tissue block method,with suitable culture condition and manipulation.There are obvious differences in the morphological characteristics and expression of certain protein between CAFs and NFs.更多还原