改良法培养鼠腹膜间皮细胞

目的 :建立一种从腹腔灌注消化液分离鼠腹膜间皮细胞的培养模型。方法 :用 0 .12 5 %的胰蛋白酶 - 0 .0 1%EDTANa2 消化液注入鼠腹腔 ,抽取腹腔灌注液进行培养 ,采用相差显微镜、电镜和免疫组化鉴定间皮细胞。结果 :分离并培养的细胞融合后在相差显微镜下呈铺路卵石状 ,纯度达 95 %以上。电镜下可见丰富的微绒毛。免疫组化鉴定角蛋白和波形蛋白阳性 ,Ⅷ因子和白细胞CD4 5抗原阴性。结论 :鼠腹膜间皮细胞培养模型建立成功 ,该模型可为进一步研究腹膜透析并纤维化的防治提供实验基础

A modified method for isolating and culturing peritoneal mesothelial cells in rats

Objective To establish a culture model of peritoneal mesothelial cells in vitro separated by infusing the digestive fluid in the rat abdominal cativity. Methods Peritoneal mesothelial cells (PMC) were digested with 0.125% trypsin 0.01% EDTA Na 2 in the abdominal cavity. We collected coelio perfusate and cultured it. The isolated cells were identified by the phase contrast microscope, electron microscope and immunohistochemical analysis. Results The confluent cells showed a uniform cobblestone like appearance under the phase contrast microscope;numerous surface microvilli could be found under the electron microscope and the purity of the cells was over 95%. Immunohistochemical studies revealed a positive staining for cytokeratin and vimentin, but a negative staining for FactorⅧ associated antigen and CD45. Conclusion The model of rat peritoneal mesothelial cells has been established successfully and it will provide an experimental basis of studying the fibrosis in peritoneal dialysis.