人脐静脉内皮细胞分离培育的改进及其鉴定【摘要】 目的 改进静脉内皮细胞原代培育的方法,提高体外培育血管内皮细胞的成功率。方法 采纳 5 号带柄一次性静脉输液针灌注消化液,分别用%、%胰蛋白酶和%胶原酶消化脐静脉内膜,比较三种酶液消化的结果,并在倒置相差显微镜下观察其形态特点,同时用免疫组织化学的方法对所得细胞进行鉴定。结果 %、%胰蛋白酶和%胶原酶的最佳消化时间分别为15min,10min,12min。倒置相差显微镜下观察内皮细胞为单层生长,鹅卵石样镶嵌排列。免疫组织化学检测表明细胞内有Ⅷ因子相关抗原。结论 胶原酶是分离培育脐静脉内皮细胞的首选消化液,且%胶原酶消化 12min 获得的细胞数量多,成活率高。 【关键词】 人脐静脉 内皮细胞 细胞培育 分离 改进 Improvement of separation ,culture and identification of human 【Abstract】 Objective To investigate the primary culture method of ehdothelial cells from human umbilical vein and improve the rate of successful culture in %,% trpsin and % collagenase were injected into the human umbilical vein by using disposable vein transfusion needle ,after which the endothelial cells were identified by morphology and immunohistochemistry under inverted The cultured cells had a cobble stone appearance with a strict monolayer growth and had Ⅷ related antigen. There are many endothelial cells after digested by %,% trpsin and % collagenase for 15 minutes,10 minutes,12 minutes The collagenase was the best choice to isolate ehdothelial cells from human umbilical vein. 【Key words】 human umbilical vein endothelial cells cell culture separation improvement 血管内皮细胞具有多种生理功能,能产生和分泌许多生物活性物质,在维持血管舒缩,抗凝血等方面起重要作用。体外血管内皮细胞的成功培育是讨论内皮细胞功能及其在各种疾病发生进展中所起作用的基础。本文报告利用健康产妇正常娩出的胎盘端游离脐带,采纳改进的灌流消化法[1]分离脐静脉内皮细胞成功传代培育并予鉴定,方法可靠,简便易行,有利于后续实验讨论的进行。 1 材料与方法 标本采集 在无菌条...