原代人脐静脉内皮细胞分离培养模型的建立  

A Model for Isolation and Culture of Primary Human Umbilical Vein Endothelial Cells

作　　者：黎胜苗[1] 罗春芬[1] 卢兰琴[2] 朱敏[3] 於林军[1] 刘昭蓉[1] 苏宝利[1] 

机构地区：[1]温州医学院附属台州医院小儿外科,317000 [2]温州医学院附属台州医院产科,317000 [3]温州医学院附属台州医院公共科研平台,317000

出　　处：《医学研究杂志》2013年第5期94-96,共3页Journal of Medical Research

基　　金：浙江省医药卫生科技计划项目(2012KYB235)

摘　　要：目的建立一套稳定的原代人脐静脉内皮细胞(HUVECs)的体外培养模型。方法新鲜人脐带标本经三通管装置胶原酶灌注法消化获取HUVECs,分别进行细胞形态学观察、Ⅷ因子相关抗原鉴定及细胞生长曲线的测定。结果从人脐静脉获取的HUVECs纯度较高,4h细胞即开始贴壁,24h基本贴壁完全,3～6天细胞生长迅速,7～8天开始融合生长,倒置相差显微镜下呈特征性的"铺路石样"镶嵌排列;传代细胞接种第1天细胞数量无明显变化,3～6天生长比较迅速,7天后进入平台期,细胞数量无明显增加;Ⅷ因子相关抗原鉴定阳性。结论采用三通管装置胶原酶灌注法可获取高纯度的HUVEC,是一种简单、实用、可推广的原代培养模型。Objective To establish a stable model for isolation and culture of primary Human umbilical vein endothelial cells （HUVECs） in vitro. Methods Fresh umbilical cords were digested by collagenase solution to obtain a single cell suspension of HUVECs. HUVECs were identified for obserration of morphological eharaeteristics, idencification of factor Ⅷ related antigen and determination of cell growth curve separately with immunoehemieal method. Results The model helped to obtain high purity HUVECs from umbilical vein. Typical ＂cobblestone＂ morphology were showed under inverted microscope, Factor Ⅷ related antigen was positive by immunostaining. Conclusion High purity of HUVECs could be obtained by collagenase solution perfusion. This model is a simple, practical and worth popularizing method of primary culture.

关 键 词：内皮细胞 脐静脉 原代培养 

分 类 号：R285.5[医药卫生—中药学]