A simple and highly effective method for the stable transduction of uncultured porcine hepatocytes using lentiviral vector

Article Article scientifique Article A simple and highly effective method for the stable transduction of uncultured porcine hepatocytes using lentiviral vector eng 2005 author Nguyen Tuan Huy 133497 author Khakhoulina Tatiana 134221 author Simmons Andrew author Morel Philippe 22337 author Trono Didier 116006 489-496 10.3727/000000005783982828 16285257 0963-6897 Gene therapy is an attractive approach for the treatment of a wide spectrum of liver diseases. Lentiviral vectors allow the stable integration of transgenes into the genome of nondividing differentiated cells including hepatocytes and could provide long-lasting expression of a therapeutic gene. To develop such approaches, preclinical studies in large animal models such as pigs are necessary to evaluate the feasibility and safety of stable lentiviral integration and long-term vector expression. In addition, effective lentivector-mediated gene transfer onto porcine hepatocytes could advance in cell-based therapies for acute liver failure. To investigate this issue, porcine hepatocytes were transduced in suspension immediately after their isolation in University of Wisconsin (UW) solution containing vitamin E. Up to 80% of hepatocytes stably expressed a GFP transgene after a single exposure to lentiviral vector coding for GFP under the control of either liver-specific or ubiquitous promoters. Moreover, porcine hepatocytes cryopreserved in UW solution containing fetal bovine serum, dimethyl sulfoxide, and vitamin E remained highly transducible with lentiviral vector after thawing. When thawed, transduced in suspension, and immediately transplanted into the spleen of immunodeficient mice, ex vivo lentivirally transgene marked xenogeneic hepatocytes were detected in murine liver. We demonstrated that porcine hepatocytes are highly susceptible to lentiviral vector and describe an easy methodology to efficiently, rapidly, and stably introduce transgenes into uncultured porcine hepatocytes. Animals Cell Separation Dose-Response Relationship, Drug Genetic Vectors Hepatocytes/cytology/transplantation Lentivirus/genetics Liver/chemistry/cytology Mice Mice, Inbred NOD Mice, SCID Swine Tissue Engineering Transduction, Genetic/methods Dewey 617 57 Département de chirurgie 222 false 79 Chirurgie viscérale 104 1329 Chirurgie viscérale (HUG) 270 Cultures cellulaires et transplantations 519 Cell transplantation 14 7 96835537-d6d4-418d-b0eb-a4c4e117de5d Article (Published version) Restricted s8.pdf 202267 application/pdf restrictedAccess Faculté de médecine / Section de médecine clinique / Département de chirurgie

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A simple and highly effective method for the stable transduction of uncultured porcine hepatocytes using lentiviral vector	Cell transplantation			2005	733	0

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Khakhoulina, Tatiana