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.
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
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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.
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