Ex Vivo Model in Cholestasis Research

  • Ruby E. H. Karsten
  • Dorenda Oosterhuis
  • Louise A. van Wijk
  • Peter OlingaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1981)


To mimic (human) cholestasis in vitro requires multiple triggers to establish a diseased phenotype. However, this is currently not simulated by existing in vitro models. Therefore, there is a high need for multicellular systems similar to the human physiology. In such an in vitro model, cell-cell interactions and intact bile canaliculi with functional bile flow should be present and preserved during long-term culture. Precision-cut liver slices represent an ex vivo tissue culture technique that replicates most of the multicellular characteristics of a whole liver in vivo. This chapter describes the preparation and culturing of (human) precision-cut liver slices. Furthermore, a protocol to use the precision-cut liver slices technique to predict drug-induced cholestatic liver injury is described.

Key words

Precision-cut liver slices ATP assay Protein assay Ex vivo model Rat bile acid mixture Human bile acid mixture Krebs Henseleit buffer Williams’ medium E University of Wisconsin Organ preservation solution 



This work was financially supported by the grants of ZonMW (114000098, 114021010, 114025003, and 114022505) and EU Horizon2020 FETOPEN-01-2016-2017 (Project number: 190634130). Prof. Dr. E.M.J. Verpoorte for editing the manuscript.


  1. 1.
    Starokozhko V, Greupink R, van de Broek P et al (2017) Rat precision-cut liver slices predict drug-induced cholestatic injury. Arch Toxicol 91:3403–3413CrossRefGoogle Scholar
  2. 2.
    Cheng Y, Woolf TF, Gan J et al (2016) In vitro model systems to investigate bile salt export pump (BSEP) activity and drug interactions: a review. Chem Biol Interact 255:23–30CrossRefGoogle Scholar
  3. 3.
    Bell CC, Hendriks DF, Moro SM et al (2016) Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease. Sci Rep 6:25187CrossRefGoogle Scholar
  4. 4.
    Hendriks DF, Fredriksson Puigvert L, Messner S et al (2016) Hepatic 3D spheroid models for the detection and study of compounds with cholestatic liability. Sci Rep 6:35434CrossRefGoogle Scholar
  5. 5.
    Lauer B, Tuschl G, Kling M et al (2009) Species-specific toxicity of diclofenac and troglitazone in primary human and rat hepatocytes. Chem Biol Interact 179:17–24CrossRefGoogle Scholar
  6. 6.
    Starokozhko V, Vatakuti S, Schievink B et al (2017) Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days. Arch Toxicol 91:2079–2092CrossRefGoogle Scholar
  7. 7.
    de Graaf IA, Olinga P, de Jager MH et al (2010) Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies. Nat Protoc 5:1540–1551CrossRefGoogle Scholar
  8. 8.
    Vatakuti S, Pennings JLA, Gore E et al (2016) Classification of cholestatic and necrotic hepatotoxicants using transcriptomics on human precision-cut liver slices. Chem Res Toxicol 29:342–351CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Ruby E. H. Karsten
    • 1
  • Dorenda Oosterhuis
    • 2
  • Louise A. van Wijk
    • 2
  • Peter Olinga
    • 2
    Email author
  1. 1.Department of Pharmaceutical AnalysisGroningen Research Institute of Pharmacy, University of GroningenGroningenThe Netherlands
  2. 2.Department of Pharmaceutical Technology and BiopharmacyGroningen Research Institute of Pharmacy, University of GroningenGroningenThe Netherlands

Personalised recommendations