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Gene delivery to pancreatic exocrine cells in vivo and in vitro
Houbracken I., Baeyens L., Ravassard P., Heimberg H., Bouwens L.
BMC Biotechnology 12, 1 (2012) 74 - http://www.hal.inserm.fr/inserm-00747861
 (23088534) 
Gene delivery to pancreatic exocrine cells in vivo and in vitro
Isabelle Houbracken () 1, Luc Baeyens2, Philippe Ravassard3, Harry Heimberg2, Luc Bouwens1
1 :  Cell Differentiation Lab
Vrije Universiteit Brussel
Diabetes Research Center, Laarbeeklaan 103, Brussels, B-1090
Belgique
2 :  Beta Cell Neogenesis Lab
Vrije Universiteit Brussel
Diabetes Research Center, Laarbeeklaan 103, Brussels, B-1090
Belgique
3 :  CRICM - Centre de Recherche de l'Institut du Cerveau et de la Moelle épinière
http://www.cricm.upmc.fr/
INSERM : U975 – Université Pierre et Marie Curie (UPMC) - Paris VI – CNRS : UMR7225
GHU Pitié-Salpêtrière 47, boulevard de l'Hôpital 75651 Paris cedex 13
France
Background
Effective gene transfer to the pancreas or to pancreatic cells has remained elusive although it is essential for studies of genetic lineage tracing and modulation of gene expression. Different transduction methods and viral vectors were tested in vitro and in vivo, in rat and mouse pancreas.
Results
For in vitro transfection/transduction of rat exocrine cells lipofection reagents, adenoviral vectors, and Mokola- and VSV-G pseudotyped lentiviral vectors were used. For in vivo transduction of mouse and rat pancreas adenoviral vectors and VSV-G lentiviral vectors were injected into the parenchymal tissue. Both lipofection of rat exocrine cell cultures and transduction with Mokola pseudotyped lentiviral vectors were inefficient and resulted in less than 4% EGFP expressing cells. Adenoviral transduction was highly efficient but its usefulness for gene delivery to rat exocrine cells in vitro was hampered by a drastic increase in cell death. In vitro transduction of rat exocrine cells was most optimal with VSV-G pseudotyped lentiviral vectors, with stable transgene expression, no significant effect on cell survival and about 40% transduced cells. In vivo, pancreatic cells could not be transduced by intra-parenchymal administration of lentiviral vectors in mouse and rat pancreas. However, a high efficiency could be obtained by adenoviral vectors, resulting in transient transduction of mainly exocrine acinar cells. Injection in immune-deficient animals diminished leukocyte infiltration and prolonged transgene expression.
Conclusions
In summary, our study remarkably demonstrates that transduction of pancreatic exocrine cells requires lentiviral vectors in vitro but adenoviral vectors in vivo.
Sciences du Vivant/Biotechnologies
Informatique/Biotechnologie
Anglais

Articles dans des revues avec comité de lecture
10.1186/1472-6750-12-74
BMC Biotechnology (BMC Biotechnol)
Publisher BioMed Central
ISSN 1472-6750 
internationale
2012
12
1
74

Lentiviral vector – Adenoviral vector – Lipofection – Gene transfer – Pancreas – Acinar cell
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