人卵巢癌微血管内皮细胞的分离培养与鉴定.doc

人卵巢癌微血管内皮细胞的培养与鉴定 徐 燕 梁志清 王 丹 （第三军医大学西南医院妇产科，重庆 400038) 【摘要】 目的建立人卵巢癌微血管内皮细胞(ovarian carcinoma-derived microvascular endothelial cells ,ODMEC)体外培养体系。方法采用胰酶、胶原酶、dNase酶联合消化法分离微血管内皮,再利用结合CD31抗体的MACS MicroBeads 免疫磁珠分选系统纯化ODMEC。光镜、流式细胞术、免疫细胞化学对所获得的ODMEC进行鉴定。结果所获ODMEC流式CD31分析细胞纯度可达99.6%表达CD34CD31、FⅧ-Rag培养中的内皮细胞生长状态良好呈现典型的铺路石征，有接触抑制可传代培养ODMEC在无明胶包被的培养池内，于第5周-6周可自发性地形成管腔样结构。 结论本研究成功建立了人ODMEC的卵巢癌血管生成研究。 【关键词】 微血管内皮细胞；卵巢癌；细胞培养；免疫磁珠 ulture and identification of microvascular endothelial cells from human ovarian carcinoma XU Yan，LIANG Zhi-qing，WANG Dan （Departemnt of Obstetrics and Gynecology,the Southwest Hospital of the Third Military Medical University,Chongqing 400038,China） 【Abstract 】 Objective To establish the method for isolating and culturing human ovarian carcinoma-derived microvascular endothelial cells (ODMECs). 基金项目：国家自然科学基金项目资助（No.；“十一五”全军科技计划青年基金项目（06Q054） 作者简介：徐燕（1979－），女，硕士研究生，主要从事妇科肿瘤方面的研究。电话 xymk__2008@163.com 通信作者：梁志清，E-mail: zhi. lzliang@gmail. com，（023王丹，E-mail:wangd717@，（023Methods The tissues of human epithelial ovarian carcinoma were digested by Trypsin,collagenase and dNase I. The cells were purified using a MACS magnetic cell sorting system with antiCD31 antibody. ODMECs were identified by light microscope，flow cytometry, immunocytochemistry. Results The ODMECs grew as gconfluentmonolayer like cobblestones and kept their features through passages. They expressed CD34,CD31, FⅧ-RAg and the purity was about 99.6 %. They bing of Ulex europaeus lectin and uptake of Acetylated-Low-Density Lipoprotein. Conclusion We have established a useful for the studies of human epithelial ovarian carcinoma. practical method for isolating ODMECs,which could be useful for the studing ideal anti-angiogenic therapies of human epithelial ovarian carcinoma. 【Key words】microvascular endothelium cell; ovarian carcinoma；cell culture；magnetic beads(tumor-derived microvascular endothelial cells ,TDMEC)具有遗传学稳定性的特点，不易获得耐药性，实际上很多基于