猪脐静脉血管内皮细胞永生化的初步研究  

Study on immortalization of swine umbilicus veins endothelial cells

作　　者：马泉荔[1] 洪海霞[1] 张彦明[1] 徐浩[1] 杨蒙[1] 苏正元[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2007年第7期43-47,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(编号:30471290)

摘　　要：为了建立稳定的血管内皮细胞系,进而为研究猪瘟病毒致病机理提供理想的细胞模型,将脂质体转染的pCIneo-hTERT质粒导入原代猪脐静脉血管内皮细胞中,经G418抗性筛选,挑取阳性克隆并扩大培养。对转染后的细胞进行形态观察,研究其增殖及生长特征,并对其端粒酶活性进行了鉴定。结果表明,转染细胞单层贴壁生长,有接触抑制性,呈铺路石状;F-ⅧAg和CD34免疫荧光鉴定阳性,具有转染前细胞特征;转染后细胞培养代数增加,传至第10代和15代转染后细胞端粒酶活性检测为阳性。说明hTERT转染可以延长猪血管内皮细胞的寿命。To establish stable vascular endothelial cell line as an ideal cell model in the research of facilitating the study on pathologic mechanism of CSFV, swine umbilical vein endothelial primary cells were transfected with the plasmid pCIneo-hTERT by liposome. Positive cells clone were selected by G418. The morphous, proliferation and growth characteristics of tansfected cell were observed and studied, including the identification of telomerase activation. The result showed that the confluent monolayer transfected cells looked like cobblestones with contact inhibition. Immunofluorescence staining showed the transfected cells Factor-Ⅷ and CD34 positive. The number of cell passage increased after transfection and the telomerase activation was positive at the 10th passage and 15th passage. The results demonstrated the transfection of hTERT gene could prolong the swine umbilical vein endothelial cell survival.

关 键 词：端粒酶催化亚基 脐静脉血管内皮细胞 猪 

分 类 号：Q813.11[生物学—生物工程]