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[1,2] 郑俊发[1,2] 曾曙光[1,2] 陈少鹏[3] 薛国初[1,2] 章锦才[1,2] 

机构地区：[1]南方医科大学附属口腔医院 [2]广东省口腔医院口腔颌面外科,广东广州510280 [3]中国科学院广州生物医药与健康研究所,广东广州510663

出　　处：《华西口腔医学杂志》2009年第6期653-656,共4页West China Journal of Stomatology

基　　金：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)

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-2mRNA和蛋白表达情况进行检测。结果原代培养的HUVECs约于24h完全贴壁,4～5d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-2mRNA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。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 immuno- cytochemistry. 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.

关 键 词：人脐静脉内皮细胞 酪氨酸激酶受体-2 肿瘤 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]