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Isolation and culture of human peritoneal mesothelial cells

Liu Hongfa;Zang Yan; Hou Fanfan; Wang Li(Department of Nephrology, Nanfang Hospital, First Military Medical University, Guangzhou, 510515)  
Objective In vitro cultured human peritoneal mesothelial cell is a useful model for studying the peritoneal function. Amodified digestiVe method was developed to separate human peritoneal mesothelial cells (HPMC) from human greater omenta(HGO). Methods HGO obtained at surgery were cut into small pieces and digested with 0.2% trypsin-0.02%EDTANa2 for 25minutes. Cell suspension was passed through a 100 mesh and cultured in RPMI-1640 containing 10% fatal calf serum. Isolated cellswere characterized by White-Giemsa staining and immunohistochemical analysis. Results The confluent cells showed a uniformcobblestone-like appearance, with a purity95%. Immunohistochemical studies showed positive staining for cytokeratin andvimentin, but negative staining for factor Ⅷ associated antigen and CD45, indicating a pure population of HPMC. ConclusionTrypsin digestion method is a simple and useful protocol for isolation of HPMC.
【Fund】: 国家自然科学基金!39470337
【CateGory Index】: R329.2
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