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Mol. Hum. Reprod. Advance Access published online on June 13, 2006
Molecular Human Reproduction, doi:10.1093/molehr/gal054
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received March 29, 2006
Revised May 8, 2006
Accepted May 16, 2006

Article

Immortalization of normal human cytotrophoblast cells by reconstitution of telomeric reverse transcriptase activity

Yan-ling Wang 1 *, Wei Qiu 1, Hui-chen Feng 2, Yu-xia Li 1, Lin-zhi Zhuang 1, Zhe Wang 1, Yu Liu 2, Jin-qiu Zhou 3, Deng-hong Zhang 3, and George S.W. Tsao 2

1 State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing
2 Cancer Biology Laboratory, Department of Anatomy, University of Hong Kong, Hong Kong
3 State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China

* To whom correspondence should be addressed.
Yan-ling Wang, E-mail: wangyl{at}ioz.ac.cn


  Abstract

Placental trophoblast cells are unique endocrine cells that play vital roles during the processes of embryonic implantation and placentation. However, research into the function of human trophoblast has been largely restrained mainly due to a lack of adequate cell models. A normal placenta-origin cytotrophoblast cell line (NPC) was previously established by our group, but these cells showed replicating senescence after 50 population doublings (PDs). In this study, the human telomerase catalytic subunit gene (htert) was transferred into B6 strain of NPC cells, and strains with reconstituted telomerase activity (B6Tert) were established. It was shown that B6Tert-1 cells produce various biomarkers of normal extravillous cytotrophoblasts during the early weeks of gestation. Meanwhile, the cell invasiveness was inhibited by transforming growth factor {beta} (TGF{beta}). However, their ability to form syncytium was relatively low when stimulated with fetal calf serum (FCS). The cells maintained normal cell growth properties and failed to elicit tumours in nude mice. They proliferated continuously with no signs of senescence until the final count at 210 PDs. The growth rate of B6Tert-1 cells was increased when compared with the parental cells, which results, at least partly, from facilitating release of the G1/S checkpoint during the cell-cycle regulation. This is the first report of immortalizing human normal cytotro-phoblast (CTB) cells by activation of telomerase activity. The cells will provide an ideal in vitro model for the study of human extravillous trophoblast (EVT) functions and consequently the mechanisms of embryonic implantation and placentation.

Keywords: G1/S checkpoint/human cytotrophoblast/immortalization/telomerase.
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