三种人脐静脉内皮细胞分离方法的比较

目的 建立人脐静脉内皮细胞（HUVECs）分离和培养的方法.方法 分别采取“灌注消化法”、“机械刮取法”和“灌注搓揉法”来分离HUVECs.通过苔盼蓝排斥法检测细胞活力并计数,进行免疫荧光法鉴定,比较这3种方法在获取的细胞数目、细胞活力和纯度方面的差异.常规进行细胞的原代培养和传代培养.取第3代培养细胞绘制细胞生长曲线,根据细胞生长特点,形态及表型特征对培养细胞进行鉴定.结果 “灌注搓揉法”所分离的细胞无论从细胞数目、细胞活力和纯度方面均优于传统的“灌注消化法”和“机械刮取法”.培养的细胞从形态、生长特点和表型鉴定证实为血管内皮细胞（ECs）.结论 “灌注搓揉法”是一种获取血管ECs成功率高,可靠的方法.

Comparison of Three Isolated Techniques on Human Umbilical Vein Endothelial Cells

Objective To establish the methods of isolation and culture of human umbilical vein endothelial cells(HUVECs).Methods HUVECs,isolated by "irrigative digestion","mechanical scrape" and "irrigative kneading" method,respectively,were used to test the cell viability and number by trypan blue exclusion and identify the phenotypes by immunofluoremetry.The differences among these methods in cell number,viability and purity were compared to identify an optimal method.The isolated HUVECs were performed routinely culture.Third-passage HUVECs were used to prepare cell growth curve and identify by morphology and phenotype.Results The "Irrigative kneading method" was superior to the traditional methods such as "Irrigative digestion method" or "Mechanical scrape method" for cell number,viability and purity.The cells isolated by these techniques were identified as HUVECs by morphology,growth features,and immunofluoremetry.Conclusion "Irrigative kneading method" would be a reliable technique to obtain HUVECs with high success rate.