Lin Shaofen, Zhou Huanjiao, Ding Yungang, Liang Xiaoling, Luo Yan, Tang Shibo
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China.
Yan Ke Xue Bao. 2010 Nov;25(2):99-102. doi: 10.3969/g.issn.1000-4432.2010.02.010.
To establish a simple and convenient method for the culture of human umbilical vein endothelial cells (HUVECs) and study its characterization in vitro.
Human Umbilical cord was isolated and digested by collagenase type I,and then it was cultured in 1.5% geltain coated dish with 10% fetal bovine serum(FBS)，heparin sodium andβ-endothelial cell growth factor(β-ECGF) in human endothelial basal growth medium．HUVECs were identified by anti-human factor Ⅷ related antigen and CD31 immunohistochemical staining．
We could successfully culture HUVECs by using this method. HUVECs attached the dish in 24 hours and the confluence was seen in 5 days. HUVECs were generally positive for anti-human factor Ⅷ related antigen and CD31．
It is a fast and effective method to successfully culture purified HUVECs in which collagenase type I was used to digest HUVECs with glass tube connected to T-tube, human endothelial basal growth medium containing 10% FBS,heparin sodium and β-ECGF in 1.5% geltain coated dish.
建立一种简便的人脐静脉内皮细胞（HUVECs）培养方法，并研究其体外特性。
分离人脐带，用Ⅰ型胶原酶消化，然后将其接种于涂有1.5%明胶的培养皿中，在含10%胎牛血清（FBS）、肝素钠和β-内皮细胞生长因子（β-ECGF）的人内皮细胞基础生长培养基中培养。通过抗人因子Ⅷ相关抗原和CD31免疫组织化学染色鉴定HUVECs。
用该方法可成功培养HUVECs。HUVECs在24小时内贴壁，5天可见汇合。HUVECs抗人因子Ⅷ相关抗原和CD31一般呈阳性。
用Ⅰ型胶原酶通过连接T管的玻璃管消化HUVECs，在涂有1.5%明胶的培养皿中，于含10%FBS、肝素钠和β-ECGF的人内皮细胞基础生长培养基中成功培养纯化的HUVECs，是一种快速有效的方法。
