腹膜透析引流液离心法和大网膜消化法原代培养人腹膜间皮细胞  

Primary Culture of Human Mesothelial Cell from Peritoneal Omenta and Effluents in Dialysis Solution Respectively

作　　者：张柯[1] 周循[2] 张浩[1] 邹莎琳[2] 凌光辉[2] 孙林[2] 刘伏友[2] 

机构地区：[1]中南大学湘雅三医院肾内科,湖南长沙410013 [2]中南大学湘雅二医院肾内科,湖南长沙410011

出　　处：《医学临床研究》2010年第9期1615-1617,1620,共4页Journal of Clinical Research

基　　金：卫生部临床重点项目[2007]353号

摘　　要：[目的]建立和改善两种人腹膜间皮原代细胞培养的方法.[方法]分别采用离心法从腹膜透析（PD）引流液中及胰蛋白酶-EDTA消化法从人的腹膜组织中分离培养人腹膜间皮细胞（HPMC）,并分别对两种原代培养的HPMC采用相差倒置显微镜和免疫组化染色对培养的细胞进行鉴定.[结果]①两种方法均在体外原代培养成活,在相差倒置显微镜下观察细胞均呈典型的铺路石样外观,免疫组化显示两种方法培养的HPMC胞浆角蛋白、波形蛋白表达阳性,提示为腹膜间皮细胞.②经大网膜消化法获得的HPMC可传至第四代,保持原有铺路石样外观,PD引流液离心法获得的HPMC传代后生长缓慢,长期接受PD后HPMC可呈现成纤维细胞样改变.[结论]成功应用离心法和消化法进行HPMC原代培养,有助于PD研究体外实验的开展.[Objective] To establish the methods of primary culture of human mesothelial cell（HPMC） from peritoneal omenta and effluents in dialysis solution. [Methods] HPMC was primarily cultured by eentrifugation from effluents in dialysis solution and trypsinase-EDTA digest from peritoneal omenta respectively, which was verified by morphology and immunohistoehemical method. [Results] HPMC was successfully primarily cultured by both the methods, which presented a typical cobble-stone appearance. Immunohistochemistry showed that HPMC kytoplasm cytokeratin and vimentin cultured by two methods were positive, which indicated peritoneal mesothelial cells. The HPMC obtained from omenta was successfully passaged. The HPMC obtained from effluents of long-term peritoneal dialysis patients presented a fibroblast appearance. [Conclusion] HPMC is successfully primarily cultured by both the methods, which benefit to investigate peritoneal dialysis in vitro.

关 键 词：腹膜透析 引流 离心法 腹膜/细胞学 间质细胞 

分 类 号：R459.5[医药卫生—治疗学]