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Reversible immortalization of rat pancreatic β cells with a novel immortalizing and tamoxifen-mediated self-recombination tricistronic vector
Wu H.-L.1; Wang Y.2; Zhang P.3; Li S.-F.1; Chen X.4; Chen Y.-K.1; Li J.-G.1; Yang S.-M.1; Su Y.-P.1; Wang J.-P.1; Chen B.1
2011-02-10
Source PublicationJournal of Biotechnology
ISSN01681656
Volume151Issue:3Pages:231-241
Abstract

Although the strategy of " Cre/LoxP-based reversible immortalization" holds great promise to overcome the cellular senescence of primary cell cultures for their further use, a secondary gene transfer for Cre expression is usually utilized to trigger the excision of the immortalizing genes in a large number of cells, thus presenting a formidable hurdle for large-scale application. We modified the strategy by utilizing a tricistronic retroviral vector pLCRSTP, in which Cre-ER, simian virus 40 large T antigen (SV40LTAg) oncogene, and a reporter gene were flanked by the same pair of LoxA sites. Five immortalized rat pancreatic β cell clones transduced with pLCRSTP, and six immortalized rat pancreatic β cell clones co-transduced with pLCRSTP and another vector encoding the human telomerase reverse transcriptase (hTERT) gene, were obtained, respectively. The Cre-ER protein could be induced to translocate from the cytoplasm to the nucleus by 4-hydroxytamoxifen to make SV40LTAg, hTERT and the Cre-ER gene itself excise without a secondary gene transfer. Our studies suggest that this system is useful to expand rat β cells and may allow for large-scale production due to its simpler manipulation. © 2010 Elsevier B.V.

KeywordCre-er/loxp Immortalization Rat Pancreatic β Cells Tricistronic
DOI10.1016/j.jbiotec.2010.12.003
URLView the original
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000287427600001
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Cited Times [WOS]:10   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
Affiliation1.Third Military Medical University
2.Daping Hospital, the Third Military Medical University
3.Chongqing Medical University
4.Universidade de Macau
Recommended Citation
GB/T 7714
Wu H.-L.,Wang Y.,Zhang P.,et al. Reversible immortalization of rat pancreatic β cells with a novel immortalizing and tamoxifen-mediated self-recombination tricistronic vector[J]. Journal of Biotechnology,2011,151(3):231-241.
APA Wu H.-L..,Wang Y..,Zhang P..,Li S.-F..,Chen X..,...&Chen B..(2011).Reversible immortalization of rat pancreatic β cells with a novel immortalizing and tamoxifen-mediated self-recombination tricistronic vector.Journal of Biotechnology,151(3),231-241.
MLA Wu H.-L.,et al."Reversible immortalization of rat pancreatic β cells with a novel immortalizing and tamoxifen-mediated self-recombination tricistronic vector".Journal of Biotechnology 151.3(2011):231-241.
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