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Human Hepatocyte Culture

  • Protocol
Cytochrome P450 Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 320))

Abstract

Primary culture of human hepatocytes is an in vitro model widely used to investigate numerous aspects of liver physiology and pathology. The technique used to isolate human hepatocytes is based on two-step collagenase perfusion. Originally performed in situ for obtaining hepatocytes from the adult rat, this technique has been adapted to the ex vivo treatment of human liver from organ donors or from lobectomy resection for medical purposes. This chapter describes experimental protocols for the isolation of hepatocytes from human liver tissue and for the preparation of short- and long-term cultures in which cells retain a differentiated phenotype for at least 1 mo. The various aspects emphasized here include the conditions for obtaining tissue, quality control of tissue for efficient perfusion, collagenase perfusion parameters, solutions for perfusion and culture media, cell substrate, cell plating, specific equipment, and safety conditions.

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Acknowledgments

We wish to thank Professors Jacques Domergue and Jean Michel Fabre (CHR Saint Eloi, Montpellier, France), Henri Joyeux (Centre Anticancéreux Val d’Aurelle, Montpellier, France), Gilles Fourtanier (CHR Rangueil, Toulouse, France), Patrice Le Treut (CHR la Conception, Marseille, France), and Jean Baulieux (CHR La Croix Rousse, Lyon, France) for providing human liver tissue, and Dr. Colin Young for editorial assistance. This work was supported in part by Synthelabo-Recherche Chilly-Mazarin, France; and Glaxo-France.

Author information

Authors and Affiliations

  1. INSERM U 632, Montpellier, France

    Lydiane Pichard, Edith Raulet & Patrick Maurel

  2. Sanofi-Aventis, Montpellier, France

    Gérard Fabre & Jean Bernard Ferrini

  3. AFSSAPS Montpellier, Vendargues, France

    Jean-Claude Ourlin

Authors
  1. Lydiane Pichard
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  2. Edith Raulet
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  3. GĂ©rard Fabre
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  4. Jean Bernard Ferrini
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  5. Jean-Claude Ourlin
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  6. Patrick Maurel
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Editor information

Editors and Affiliations

  1. School of Biological Sciences, Queen Mary University of London, London, UK

    Ian R. Phillips

  2. Department of Biochemistry and Molecular Biology, University College London, London, UK

    Elizabeth A. Shephard

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© 2006 Humana Press Inc.

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Pichard, L., Raulet, E., Fabre, G., Ferrini, J.B., Ourlin, JC., Maurel, P. (2006). Human Hepatocyte Culture. In: Phillips, I.R., Shephard, E.A. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology, vol 320. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-998-2:283

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  • DOI: https://doi.org/10.1385/1-59259-998-2:283

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-58829-441-8

  • Online ISBN: 978-1-59259-998-1

  • eBook Packages: Springer Protocols

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