原代人腹膜间皮细胞培养的方法学探讨

Exploration on methodology of human omentum access to primary HPMCs

目的建立简便有效的获取人腹膜间皮细胞(HPMCs)的方法。方法采用胰蛋白酶-EDTA(0.25%胰蛋白酶-0.02%EDTA-Na2)组织消化法,从人的大网膜中分离出HPMCs;分别从形态学、免疫组化鉴定分离出的细胞。结果分离出的细胞在倒置相差显微镜下早期呈多形性、拉伸状;后期呈多边形、"铺路石状"。细胞表面标志物鉴定显示角蛋白、波形蛋白抗原阳性,第Ⅷ因子相关抗原、白细胞CD45抗原阴性,证实分离培养的确是HPMCs。结论胰蛋白酶-EDTA组织消化法是一种简便有效的分离HPMCs的方法。

Objective It is to establish a simple and effective human primary culture model of peritoneal mesothelial cells （HPMCs）. Methods Trypsin - EDTA （0.25% trypsin - 0.02% EDTA Na2 ） organization of digestion was used to isolated HPMCs from human omentum. The isolated cells were respectively identified acoording to morphology and immunohistochemistry. Results Cells isolated showed multiple deformation and stretching like under the inverted phase contrast microscope in early stage, and polygonal shape,＂cobblestone like＂ in late stage. Cell surface markers identified keratin, vimentin antigen positive;Factor VⅢ related antigen, leukocyte CD45 antigen negative,which confirmed that the cells isolated and cultured were HPMCs indeed. Conclusion Trypsin - EDTA tissue digestion method is a simple and effective method for separating HPMCs.