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0.125%的胰蛋白酶－0.01％EDTA Na2消化液注入鼠腹腔，抽取腹腔灌注液进行培养，采用相差`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CD45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 Na2 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 Factor VIII 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.