title: Prolonged expansion of human nucleus pulposus cells expressing human telomerase reverse transcriptase mediated by lentiviral vector.
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.
authors: Department of Orthopaedic Surgery, Navy General Hospital, No.6 Fu-cheng Road, Beijing, 100048, PR China.
authors: Jianhong
authors: J
authors: Wu
authors: Deli
authors: D
authors: Wang
authors: Dike
authors: D
authors: Ruan
authors: Qing
authors: Q
authors: He
authors: Yan
authors: Y
authors: Zhang
authors: Chaofeng
authors: C
authors: Wang
authors: Hongkui
authors: H
authors: Xin
authors: Cheng
authors: C
authors: Xu
authors: Yue
authors: Y
authors: Liu
mesh_terms: D053674:Aggrecans / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D024043:Collagen Type II / Q000235:genetics; D004789:Enzyme Activation / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology* / Q000502:physiology*; D055959:Intervertebral Disc Degeneration / Q000235:genetics / Q000628:therapy; D059785:Karyotype; D016086:Lentivirus / Q000235:genetics*; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D016615:Telomere / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: C508319:ACAN protein, human; D053674:Aggrecans; D024043:Collagen Type II; C509186:TERT protein, human; D019098:Telomerase
keywords: degenerative disc diseases; gene therapy; human nucleus pulposus cells; human telomerase reverse transcriptase; lentiviral vector; nucleus pulposus cells; telomerase