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Int J Pharm. 2017 May 30;524(1-2):268-278. doi: 10.1016/j.ijpharm.2017.03.080. Epub 2017 Mar 30.

Efficient transfection of Xenobiotic Responsive Element-biosensor plasmid using diether lipid and phosphatidylcholine liposomes in differentiated HepaRG cells.

Author information

1
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.
2
INSERM, INRA, Univ Rennes 1, Univ Bretagne Loire, Nutrition Metabolisms and Cancer (NuMeCan), Plateforme SynNanoVect, Biogenouest, Rennes, France.
3
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.
4
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.
5
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.

Abstract

In this study, we evaluated cationic liposomes prepared from diether-NH2 and egg phosphatidylcholine (EPC) for in vitro gene delivery. The impact of the lipid composition, i.e. the EPC and Diether-NH2 molar 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.

KEYWORDS:

Archaeolipids; Biosensor plasmid; Diether lipids; HepaRG cells; Liposomes; Transfection

PMID:
28365389
DOI:
10.1016/j.ijpharm.2017.03.080
[Indexed for MEDLINE]
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