人腹膜间皮细胞的原代培养及传代

目的探讨建立一种有效的HPMC体外原代培养及传代的方法。方法用0.25%的胰蛋白酶0.02%EDTANa2消化腹部择期手术者大网膜组织,细胞悬液经100目筛网滤过后进行培养。采用形态学、免疫组织化学（ABC法）、电镜等鉴定细胞。结果分离培养的细胞光镜下呈铺路样鹅卵石状,免疫组化鉴定显示角蛋白、波形蛋白抗原阳性,第Ⅷ因子、白细胞CD45抗原阴性,电镜下可见细胞表面的微绒毛,证明培养的细胞的为HPMC。结论胰蛋白酶消化法是一种简单实用的培养HPMC的方法 。

Primary Culture and Passage of Human Peritoneal Mesothelial cells

Objective To establish a effective method of primary culture and passage of human peritoneal mesothelial cells in vitro.Methods Human Omental specimens were obtained from patients undergoing elective abdominal surgery and digested with 0.25% trypsin-0.02%EDTANa2 for 30 minutes.Cell suspension was passed through a 100 mesh and cultured in RPMI-1640 containing 20% fatal calf serum.Isolated cells were characterized by morphology,immunohistochemical analysis and electron microscopy.Results The confluent cells showed a uniform cobblestone-like appearance.Immunohistochemical studies showed positive staining of cytokeratin-8,cytokeratin-18 and vimentin,but negative staining of factor Ⅷ associated antigen and CD45.Electron microscopy showed numerous microvilli on the upper surface of the cultured cells,indicating a pure population of HPMC.Conclusion Trypsin digestion method is a simple and useful protocol for culture of HPMC.