Immortalization of swine umbilical vein endothelial cells with human telomerase reverse transcriptase.
Swine endothelial cells are commonly used as an in vitro model for studying features of the blood-brain barrier and some hemorrhagic diseases. However, primary cultures of swine cells have finite lifespans. To establish immortalized swine umbilical vein endothelial cells (SUVECs) using human telomerase reverse transcriptase (hTERT), the plasmid pCI-neo-hTERT was transfected into SUVECs by lipofection. Clones were selected for G418 resistance, and positive clones were amplified. One of the clones was cultured for up to 50 passages. Factor VIII-related antigen and CD34 were detected. The immortalized cells shared the properties of normal cells, such as contact inhibition, serum requirement and anchorage dependence. Karyotype analysis revealed that the immortalized cells were in the diploid range. In addition, both in vivo and in vitro assays of tumorigenicity showed no neoplastic transformation. Furthermore, NO, PGI2, and ET-1 concentrations in the transfected cells were normal. These results suggest that the SUVECs immortalized by hTERT retain their original characteristics.
Hong
Hai Xia
HX
College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi 712100, China.
Zhang
Yan Ming
YM
Xu
Hao
H
Su
Zheng Yuan
ZY
Sun
Pei
P
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D042783:Endothelial Cells / Q000166:cytology*; D019332:Endothelin-1 / Q000378:metabolism; D011464:Epoprostenol / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D009569:Nitric Oxide / Q000378:metabolism; D034421:Sus scrofa; D019098:Telomerase / Q000378:metabolism*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D019332:Endothelin-1; D009569:Nitric Oxide; D011464:Epoprostenol; D019098:Telomerase