目的建立人卵巢癌微血管内皮细胞(ovarian carcinoma-derived microvascular endothelial cells,ODMECs)体外培养体系。方法采用胶原酶、胰酶联合消化法分离微血管内皮,经pereoll梯度密度离心纯化。光镜、电镜、流式细胞术、免疫细胞化学对所获得的ODMECs进行鉴定。结果所获0DMECs流式分析有内皮标志物CD34/VEGF—R2表达;免疫荧光FⅧ一RAg阳性;电镜下见细胞多核、有丰富的微丝、weibel—Palade小体等;培养中的内皮细胞生长状态良好、呈现典型的铺路石征,可传代培养。结论本研究建立了人0DMECs体外培养体系,对了解卵巢癌血管内皮的异质性有重要价值,为抗卵巢癌血管生成的研究奠定了基础。
Objective To establish the method for isolating and culturing human ovarian carcinoma-derived micro- vascular endothelial cells (ODMECSs). Methods The tissues of human epithelial ovarian carcinoma were finely minced with scissors, then digested by Trypsin and collagenase. The cells were purified using percoll. ODMECSs were identified by light microscope,electron microscopy, flow cytometry and immunocytochemistry. Results The ODMECSs grew as confluent monolayer like cobblestones. They expressed F~,~-RAg/CD34/VEGF-R2. Weibel-Palade bodies could be found by electron microscopy. Conclusion We have established a useful method for isolating ODMECSs, which could be useful for the studies of human epithelial ovarian carcinoma.