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Prolonged expansion of human nucleus pulposus cells expressing human telomerase reverse transcriptase mediated by lentiviral vector.

Authors :
Wu, Jianhong
Wang, Deli
Ruan, Dike
He, Qing
Zhang, Yan
Wang, Chaofeng
Xin, Hongkui
Xu, Cheng
Liu, Yue
Source :
Journal of Orthopaedic Research. Jan2014, Vol. 32 Issue 1, p159-166. 8p.
Publication Year :
2014

Abstract

ABSTRACT Human degenerative disc disease (DDD) is characterized by progressive loss of human nucleus pulposus (HNP) cells and extracellular matrix, in which the massive deposition are secreted by HNP cells. Cell therapy to supplement HNP cells to degenerated discs has been thought to be a promising strategy to treat DDD. However, obtaining a large quality of fully functional HNP cells has been severely hampered by limited proliferation capacity of HNP cells in vitro. Previous studies have used lipofectamine or recombinant adeno-associated viral (rAAV) vectors to deliver human telomerase reverse transcriptase (hTERT) into ovine or HNP cells to prolong the activity of nucleus pulposus cells with limited success. Here we developed a lentiviral vector bearing both hTERT and a gene encoding green fluorescence protein (L-hTERT/EGFP). This vector efficiently mediated both hTERT and EGFP into freshly isolated HNP cells. The expressions of both transgenes in L-hTERT/EGFP transduced HNP cells were detected up to day 210 post viral infection, which was twice as long as rAAV vector did. Furthermore, we observed restored telomerase activity, maintained telomere length, delayed cell senescence, and increased cell proliferation rate in those L-hTERT/EGFP transduced HNP cells. Our study suggests that lentiviral vector might be a useful gene delivery vehicle for HNP cell therapy to treat DDD. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:159-166, 2014. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
07360266
Volume :
32
Issue :
1
Database :
Academic Search Index
Journal :
Journal of Orthopaedic Research
Publication Type :
Academic Journal
Accession number :
92037907
Full Text :
https://doi.org/10.1002/jor.22474