人肾小球微血管内皮细胞体外培养方法和鉴定的实验研究

Isolation and identification of microvascular endothelial cells of human glomerulus

目的探索人肾小球微血管内皮细胞的体外培养方法和鉴定。方法先用梯度筛网法获得高纯度肾微血管球,采用胶原酶消化法从肾微血管球内获取内皮细胞,再用单克隆增殖法进行纯化、体外培养;采用免疫组化法测定Ⅷ因子相关抗原,并在透射电镜下观察Weibel-Palade小体进行鉴定。结果体外培养出高纯度人肾小球微血管内皮细胞并传代培养12代。该细胞Ⅷ因子相关抗原阳性、细胞浆中可见Weibel-Palade小体。结论本实验成功培养出人肾小球微血管内皮细胞,并予以传代。

Objective To establish methods of culture and identification of microvascular endothelial cells of human glomerulus in vitro. Methods Two-step gradient sieve method was used. Glomerulus of relatively higher purity was obtained firstly, then renal microvascular endothelial cells were cultured in vitro after collagenase enzyme digestion, and pure renal microvascular endothelial cell population were finally obtained by monoclonal proliferation. The renal microvascular endothelial cells were further identified by factor Ⅷ-related antigen using immunohistochemical staining as well as the observation of cytoplasmic Weibel-Palade bodies with the transmission electron microscope. Results The highly purified human glomerulus microvascular endothelial cells were cultured in vitro with twelve passages. Factor Ⅷ-related antigen, and Weibel-Palade bodies were observed immunohistochemically by the transmission electron microscope. Conclusion Microvascular endothelial cells of human glomerulus can be cultured successfully in vitro by the method reported in this study.