A first experience of transduction for differentiated HepaRG cells using lentiviral technology.

Autor: Pivert A; Laboratory of Virology, University Hospital & LUNAM University and HIFIH laboratory, UPRES EA 3859, SFR 4208, Angers, France. adpivert@chu-angers.fr., Lefeuvre C; Laboratory of Virology, University Hospital & LUNAM University and HIFIH laboratory, UPRES EA 3859, SFR 4208, Angers, France., Tran CT; Laboratory of Virology, University Hospital & LUNAM University and HIFIH laboratory, UPRES EA 3859, SFR 4208, Angers, France., Baillou C; Sorbonne Université, Inserm, CNRS, Centre d'Immunologie et Maladies Infectieuses (CIMI-Paris), Paris, France., Durantel D; Cancer Research Center of Lyon (CRCL), INSERM, U1052, CNRS, University of Lyon, UMR_5286, LabEx DEVweCAN, Lyon, France., Le Guillou-Guillemette H; Laboratory of Virology, University Hospital & LUNAM University and HIFIH laboratory, UPRES EA 3859, SFR 4208, Angers, France., Lemoine FM; Sorbonne Université, Inserm, CNRS, Centre d'Immunologie et Maladies Infectieuses (CIMI-Paris), Paris, France., Lunel-Fabiani F; Laboratory of Virology, University Hospital & LUNAM University and HIFIH laboratory, UPRES EA 3859, SFR 4208, Angers, France., Ducancelle A; Laboratory of Virology, University Hospital & LUNAM University and HIFIH laboratory, UPRES EA 3859, SFR 4208, Angers, France.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2019 Sep 09; Vol. 9 (1), pp. 12910. Date of Electronic Publication: 2019 Sep 09.
DOI: 10.1038/s41598-019-49402-8
Abstrakt: Currently, there is a lack of systems for studying the role of hepatitis B viral proteins, such as HBeAg and HBcAg, on liver injury. It is necessary to develop an original tool in order to clarify the role of these viral proteins in hepatic stellate cell activation, and to understand the molecular mechanisms of liver injury. HepaRG are the most reliable hepatocyte-like cells for studying liver functions or disorders. In this paper, we demonstrate that the transduction of differentiated HepaRG (dHepaRG) cells can be performed successfully using lentiviral particles. The production of a functional Green Fluorescent Protein (GFP) assessed by Fluorescence Activated Cell Sorting and fluorescence microscopy is up to 16% of GFP positive cells using a multiplicity of infection (MOI) of 2.4. We demonstrate that this technology can allow the stable expression of GFP during the long lifecycle of the cell (up to four weeks after the cell's passage). With this innovative tool, we aim to express viral proteins such as HBeAg or HBcAg in dHepaRG cells. The preliminary results of this work shows that HBeAg can be efficiently produced in dHepaRG cells and that increased MOI allows a better production of this protein. Our future objective will be to study the role of HBc and HBe proteins on the induction of hepatic fibrosis.
Databáze: MEDLINE
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