人脐静脉内皮细胞的原代培养及鉴定  

Culture and identification of human umbilical vein endothelial cells

作　　者：付伟[1] 李慧敏[1] 苏建华[1] 缪珩[1] 鲁一兵[1] 

机构地区：[1]南京医科大学第二附属医院内分泌科,江苏南京210011

出　　处：《南京医科大学学报（自然科学版）》2009年第1期89-91,共3页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省卫生厅面上项目(H200514)

摘　　要：目的:体外进行人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)原代培养及鉴定。方法:用0.1%Ⅱ型胶原酶消化法收集HUVECs,加入含20%胎牛血清的M199培养基在37℃、5%CO2孵箱中培养,用Tryple express进行消化传代培养。采用细胞形态学观察、流式细胞仪测定CD34鉴定细胞。结果:原代培养的HUVECs在24h完全贴壁,在3～5天融合成片,呈现单层铺路石样改变。流式细胞仪测定内皮细胞纯度为75.52%。结论:Ⅱ胶原酶及Tryple express的灵活使用可获得大量连续传代培养的HUVECs。Objective：To culture and identify endothelial cells from human umbilical vein endothelial cells（HUVECs） in vitro. Methods：HUVECs were obtained using 0.1% collagenase type Ⅱ by the technique of irrigative digestion, and were cultured with M199 which contained fetal calf serum in the carbon dioxide incubator,and then were passaged using tryple express digestion. The culture cells were identified by morphology and flow cytometry detecting CD34. Results：HUVECs could adhere to the plate completely after 24 hours and got a monolayer 3-5 days later. The culture cells were HUVECs confirmed by morphology and flow cytometry. Conclusion：Using collagenase type Ⅱ and tryple express could obtain a large number of HUVECs in continuous passage culture.

关 键 词：人脐静脉内皮细胞 培养 鉴定 CD34 

分 类 号：R331[医药卫生—人体生理学]