人脐静脉内皮细胞的分离培养和鉴定及酪氨酸激酶受体-2在其中的表达
Culture and identify the human umbilical vein endothelial cells and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains in the cells
作者：吴世卿 郑俊发 曾曙光 陈少鹏 薛国初 章锦才
Author：WU Shiqing1, ZHENG Jun-fa1, ZENG Shu-guang1, CHEN Shao-peng2, XUE Guo-chu1, ZHANG Jin-cai1
收稿日期：2009-12-25 年卷（期）页码：2009,27(06):653-653-656
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：人脐静脉内皮细胞,酪氨酸激酶受体-2,肿瘤,
Key words：human umbilical vein endothelial cells,tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains,tumor,
基金项目：
广东省科技计划基金资助项目（2008B030301190）；广东省科研基金资助项目（A2006115）
中文摘要
目的分离和培养人脐静脉内皮细胞（HUVECs），对其进行鉴定，并探讨酪氨酸激酶受体-2（Tie-2）基因在HUVECs中的表达情况。方法用胰蛋白酶消化、分离HUVECs并对其进行培养，根据细胞生长特点、形态特征和Ⅷ因子免疫荧光组化技术对细胞进行鉴定。分别采用逆转录-聚合酶链反应（RT-PCR）和SABC免疫细胞化学染色对HUVECs中Tie-2 mRNA和蛋白表达情况进行检测。结果原代培养的HUVECs约于24 h完全贴壁，4～5 d后融合成单层铺路石样结构。Ⅷ因子免疫荧光组化法证实细胞是HUVECs。RT-PCR检测到HUVECs中Tie-2 mRNA条带明显，SABC免疫细胞组化法检测到Tie-2蛋白在HUVECs胞浆、核膜中呈强阳性表达，阳性率为85％以上。结论胰蛋白酶灌流消化法可获得高纯度的HUVECs，Tie-2在HUVECs中呈强阳性表达。
英文摘要
Objective To study the cultural method and identification of human umbilical vein endothelial cells （HUVECs）, and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains（Tie-2） in HUVECs. Methods HUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by Ⅷ monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction （RT-PCR） and SABC immunocytochemistry. Results HUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85％. Conclusion Highly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.
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