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Titolo:
Reversible immortalization of human primary cells by lentivector-mediated transfer of specific genes
Autore:
Salmon, P; Oberholzer, J; Occhiodoro, T; Morel, P; Lou, JN; Trono, D;
Indirizzi:
CMU, Dept Genet & Microbiol, Fac Med, CH-1211 Geneva 4, Switzerland CMU Geneva Switzerland 4 crobiol, Fac Med, CH-1211 Geneva 4, Switzerland CMU, Fac Med, Div Surg Res, CH-1211 Geneva, Switzerland CMU Geneva Switzerland CH-1211 Div Surg Res, CH-1211 Geneva, Switzerland
Titolo Testata:
MOLECULAR THERAPY
fascicolo: 4, volume: 2, anno: 2000,
pagine: 404 - 414
SICI:
1525-0016(200010)2:4<404:RIOHPC>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
LARGE-T-ANTIGEN; SINUSOIDAL ENDOTHELIAL-CELLS; SITE-SPECIFIC RECOMBINATION; HUMAN-DIPLOID FIBROBLASTS; RAT EMBRYO FIBROBLASTS; MAMMALIAN-CELLS; LENTIVIRAL VECTOR; SIMIAN VIRUS-40; TRANSGENIC MICE; IN-VIVO;
Keywords:
HIV vectors; lentiviral vectors; conditional immortalization; reversible immortalization; gene therapy; gene transfer; LoxP-Cre; SV40 large T; Bmi-1; telomerase; pancreatic beta cell; insulin;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
52
Recensione:
Indirizzi per estratti:
Indirizzo: Trono, D CMU, Dept Genet & Microbiol, Fac Med, 1 Rue Michel Servet, CH-1211 Geneva 4, Switzerland CMU 1 Rue Michel Servet Geneva Switzerland 4 neva 4, Switzerland
Citazione:
P. Salmon et al., "Reversible immortalization of human primary cells by lentivector-mediated transfer of specific genes", MOL THER, 2(4), 2000, pp. 404-414

Abstract

We exploited the ability of lentiviral vectors to govern the stable transduction of cells irrespective of their cycling status to induce the reversible immortalization of human primary cells. First, bicistronic HIV-derived lentiviral vectors expressing GFP- and the HSV1 thymidine kinase and containing the LoxP sequence in their LTR (HLox) were used to transduce HeLa cells. Cre expression led to efficient proviral deletion, and unexcised cells could be eliminated by ganciclovir treatment. A human liver biopsy was then exposed to a combination of HLox vectors that harbored either the SV40 largeT (TAg) or the human telomerase (hTERT) DNAs in place of GFP. This led to the isolation of liver sinusoidal endothelial cell (LSEC) clones that exhibited an immortalized phenotype while retaining most of the features of primary hLSEC. Complete growth arrest of these cells was observed in 2 days of Cre expression, and the resulting stationary culture could be kept for at least 2 weeks. Transduction of human adult pancreatic islets with HLox vectors coding for Tag and Bmi-1also induced the proliferation of insulin-positive cells. These results indicate that lentivectors can be used to mediate the reversible immortalization of primary nondividing cells and should allowfor the production of large supplies of a wide variety of human cells for both therapeutic and research purposes.

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Documento generato il 14/07/20 alle ore 05:21:11