title: Efficient transfection of Xenobiotic Responsive Element-biosensor plasmid using diether lipid and phosphatidylcholine liposomes in differentiated HepaRG cells.
abstract: In this study, we evaluated cationic liposomes prepared from diether-NH2and egg phosphatidylcholine (EPC) for in vitro gene delivery. The impact of the lipid composition, i.e. the EPC and Diether-NH2molar ratio, on in vitro transfection efficiency and cytotoxicity was investigated using the human HEK293T and hepatoma HepaRG cells known to be permissive and poorly permissive cells for liposome-mediated gene transfer, respectively. Here, we report that EPC/Diether-NH2-based liposomes enabled a very efficient transfection with low cytotoxicity compared to commercial transfection reagents in both HEK293T and proliferating progenitor HepaRG cells. Taking advantage of these non-toxic EPC/Diether-NH2-based liposomes, we developed a method to efficiently transfect differentiated hepatocyte-like HepaRG cells and a biosensor plasmid containing a Xenobiotic Responsive Element and a minimal promoter driving the transcription of the luciferase reporter gene. We demonstrated that the luciferase activity was induced by a canonical inducer of cytochrome P450 genes, the benzo[a]pyrene, and two environmental contaminants, the fluoranthene, a polycyclic aromatic hydrocarbon, and the endosulfan, an organochlorine insecticide, known to induce toxicity and genotoxicity in differentiated HepaRG cells. In conclusion, we established a new efficient lipofection-mediated gene transfer in hepatocyte-like HepaRG cells opening new perspectives in drug evaluation relying on xenobiotic inducible biosensor plasmids.
authors.affiliation: Ecole Nationale Supérieure de Chimie de Rennes, Institut des Sciences Chimiques de Rennes, UMR 6226 CNRS, Plateforme SynNanoVect, Biogenouest, 11 allée de Beaulieu, CS 50837, 35708 Rennes Cedex 7, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Demazeau
authors.affiliation: INSERM, INRA, Univ Rennes 1, Univ Bretagne Loire, Nutrition Metabolisms and Cancer (NuMeCan), Plateforme SynNanoVect, Biogenouest, Rennes, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Quesnot
authors.affiliation: Ecole Nationale Supérieure de Chimie de Rennes, Institut des Sciences Chimiques de Rennes, UMR 6226 CNRS, Plateforme SynNanoVect, Biogenouest, 11 allée de Beaulieu, CS 50837, 35708 Rennes Cedex 7, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ripoche
authors.affiliation: INSERM, INRA, Univ Rennes 1, Univ Bretagne Loire, Nutrition Metabolisms and Cancer (NuMeCan), Plateforme SynNanoVect, Biogenouest, Rennes, France.
authors.forename: Claudine
authors.initials: C
authors.lastname: Rauch
authors.affiliation: Ecole Nationale Supérieure de Chimie de Rennes, Institut des Sciences Chimiques de Rennes, UMR 6226 CNRS, Plateforme SynNanoVect, Biogenouest, 11 allée de Beaulieu, CS 50837, 35708 Rennes Cedex 7, France.
authors.forename: Jelena
authors.initials: J
authors.lastname: Jeftić
authors.affiliation: INSERM, INRA, Univ Rennes 1, Univ Bretagne Loire, Nutrition Metabolisms and Cancer (NuMeCan), Plateforme SynNanoVect, Biogenouest, Rennes, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Morel
authors.affiliation: Institut des Sciences Chimiques de Rennes, UMR 6226 CNRS, Université de Rennes 1, Campus de Beaulieu, 263 Avenue du Général Leclerc, F-35042 Rennes Cedex, France. Electronic address: fabienne.gauffre@univ-rennes1.fr.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Gauffre
authors.affiliation: Ecole Nationale Supérieure de Chimie de Rennes, Institut des Sciences Chimiques de Rennes, UMR 6226 CNRS, Plateforme SynNanoVect, Biogenouest, 11 allée de Beaulieu, CS 50837, 35708 Rennes Cedex 7, France. Electronic address: thierry.benvegnu@ensc-rennes.fr.
authors.forename: Thierry
authors.initials: T
authors.lastname: Benvegnu
authors.affiliation: INSERM, INRA, Univ Rennes 1, Univ Bretagne Loire, Nutrition Metabolisms and Cancer (NuMeCan), Plateforme SynNanoVect, Biogenouest, Rennes, France. Electronic address: pascal.loyer@univ-rennes1.fr.
authors.forename: Pascal
authors.initials: P
authors.lastname: Loyer
mesh_terms: D015374:Biosensing Techniques; D002412:Cations; D057809:HEK293 Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry*; D010957:Plasmids; D014162:Transfection
chemical_list: D002412:Cations; D008081:Liposomes; D010713:Phosphatidylcholines
keywords: Archaeolipids; Biosensor plasmid; Diether lipids; HepaRG cells; Liposomes; Transfection