Document Detail


Immortalization of normal human cytotrophoblast cells by reconstitution of telomeric reverse transcriptase activity.
MedLine Citation:
PMID:  16772430     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 (TGFbeta). 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 cytotrophoblast (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.
Authors:
Yan-ling Wang; Wei Qiu; Hui-chen Feng; Yu-xia Li; Lin-zhi Zhuang; Zhe Wang; Yu Liu; Jin-qiu Zhou; Deng-hong Zhang; George S W Tsao
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-06-13
Journal Detail:
Title:  Molecular human reproduction     Volume:  12     ISSN:  1360-9947     ISO Abbreviation:  Mol. Hum. Reprod.     Publication Date:  2006 Jul 
Date Detail:
Created Date:  2006-07-14     Completed Date:  2006-10-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9513710     Medline TA:  Mol Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  451-60     Citation Subset:  IM    
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China. wangyl@ioz.ac.cn
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Northern
Blotting, Western
Cell Line
DNA-Binding Proteins / genetics*,  metabolism
Female
Humans
Immunohistochemistry
Karyotyping
Keratins / analysis
Mice
Mice, Inbred BALB C
Mice, Nude
Microscopy, Phase-Contrast / methods
Neoplasms, Experimental / enzymology,  genetics,  pathology
Pregnancy
RNA, Messenger / genetics,  metabolism
Retinoblastoma Protein / genetics,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
Telomerase / genetics*,  metabolism
Telomere / genetics,  metabolism*
Transfection
Trophoblasts / cytology,  metabolism*
Tumor Suppressor Protein p53 / genetics,  metabolism
Vimentin / analysis
Chemical
Reg. No./Substance:
0/DNA-Binding Proteins; 0/RNA, Messenger; 0/Retinoblastoma Protein; 0/Tumor Suppressor Protein p53; 0/Vimentin; 68238-35-7/Keratins; EC 2.7.7.49/Telomerase

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