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Setup and Use of HepaRG Cells in Cholestasis Research

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Experimental Cholestasis Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1981))

Abstract

Since HepaRG cells can differentiate into well-polarized mature hepatocyte-like cells that synthesize, conjugate, and secrete bile acids, they represent an appropriate surrogate to primary human hepatocytes for investigations on drug-induced cholestasis mechanisms. In this chapter, culture conditions for obtaining HepaRG hepatocytes and the main methods used to detect cholestatic potential of drugs are described. Assays for evaluation of bile canaliculi dynamics and morphology are mainly based on time-lapse and phase-contrast microscopy analysis. Bile acid uptake, trafficking, and efflux are investigated using fluorescent probes. Individual bile acids are quantified in both culture media and cell layers by high-pressure liquid chromatography/tandem mass spectrometry. Preferential cellular accumulation of toxic hydrophobic bile acids is easily evidenced when exogenous primary and secondary bile acids are added to the culture medium.

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Acknowledgments

We are grateful to all the postdoctoral fellows and PhD students, in particular Dr. Ahmad Sharanek and Dr. Audrey Burban, who have contributed to our cholestasis project. We would also like to thank Dr. Remy Le Guevel from the ImPACell platform (Biosit, University of Rennes 1) for his help with imaging analyses. This work was mostly supported by the European Community through the Innovative Medicines Initiative Joint Undertaking MIP-DILI project (grant agreement number 115336), resources of which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/20072013) and EFPIA companies in kind contribution.

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Authors and Affiliations

  1. INSERM U1241, NuMeCan, Université de Rennes 1, Rennes, France

    Christiane Guguen-Guillouzo & André Guillouzo

Authors
  1. Christiane Guguen-Guillouzo
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  2. André Guillouzo
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Correspondence to André Guillouzo .

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Editors and Affiliations

  1. Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Brussels, Belgium

    Mathieu Vinken

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Guguen-Guillouzo, C., Guillouzo, A. (2019). Setup and Use of HepaRG Cells in Cholestasis Research. In: Vinken, M. (eds) Experimental Cholestasis Research. Methods in Molecular Biology, vol 1981. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9420-5_19

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  • DOI: https://doi.org/10.1007/978-1-4939-9420-5_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9419-9

  • Online ISBN: 978-1-4939-9420-5

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