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Received: 2004-09-06

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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.7 P.631~636

http://doi.org/10.1631/jzus.2005.B0631


Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen


Author(s):  BIAN Chang, ZHAO Kui, TONG Guo-xin, ZHU Yong-liang, CHEN Peng

Affiliation(s):  Department of Cardiology, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; more

Corresponding email(s):   skyoutwin@yahoo.com.cn

Key Words:  Endothelial cell, Telomerase activity, Immortalization


BIAN Chang, ZHAO Kui, TONG Guo-xin, ZHU Yong-liang, CHEN Peng. Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen[J]. Journal of Zhejiang University Science B, 2005, 6(7): 631~636.

@article{title="Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen",
author="BIAN Chang, ZHAO Kui, TONG Guo-xin, ZHU Yong-liang, CHEN Peng",
journal="Journal of Zhejiang University Science B",
volume="6",
number="7",
pages="631~636",
year="2005",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.B0631"
}

%0 Journal Article
%T Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen
%A BIAN Chang
%A ZHAO Kui
%A TONG Guo-xin
%A ZHU Yong-liang
%A CHEN Peng
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 7
%P 631~636
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B0631

TY - JOUR
T1 - Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen
A1 - BIAN Chang
A1 - ZHAO Kui
A1 - TONG Guo-xin
A1 - ZHU Yong-liang
A1 - CHEN Peng
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 7
SP - 631
EP - 636
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B0631


Abstract: 
Objective: To establish normally conditionally-immortalized human umbilical vein endothelial cells (HUVECs) by ectopic expression of the human telomerase catalytic enzyme (hTERT) and simian virus 40 large T (SV40 LT) antigen. Methods: Primary HUVECs were transfected with recombinant retrovirus containing hTERT or SV40 LT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. endothelial cell biomarkers were confirmed by examination. Results: The morphological phenotype of the transfected cells was similar to the non-transfected cells. Von Willebrand factor, hTERT and SV40 LT could be detected in transfected HUVECs. Moreover, higher telomerase activity in transfected cells was maintained for over 50 population doublings compared with only low level of endogenous telomerase transiently at early population doublings in primary HUVECs. When exposed to TNF-α (tumor necrosis factor-α), the expression of E-selectin in transfected cells was significantly up-regulated, but no alteration of endothelial lipase was found. Conclusion: Ectopic coexpression of hTERT and SV40 LT can effectively immortalize HUVECs without tumorigenicity in vitro. Immortalized HUVECs may be an ideal target of further molecular function studies.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

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