title: Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days.
abstract: Human precision-cut liver slices (hPCLS) are a valuable ex vivo model that can be used in acute toxicity studies. However, a rapid decline in metabolic enzyme activity limits their use in studies that require a prolonged xenobiotic exposure. The aim of the study was to extend the viability and function of hPCLS to 5 days of incubation. hPCLS were incubated in two media developed for long-term culture of hepatocytes, RegeneMed®, and Cellartis®, and in the standard medium WME. Maintenance of phase I and II metabolism was studied both on gene expression as well as functional level using a mixture of CYP isoform-specific substrates. Albumin synthesis, morphological integrity, and glycogen storage was assessed, and gene expression was studied by transcriptomic analysis using microarrays with a focus on genes involved in drug metabolism, transport and toxicity. The data show that hPCLS retain their viability and functionality during 5 days of incubation in Cellartis®medium. Albumin synthesis as well as the activity and gene expression of phase I and II metabolic enzymes did not decline during 120-h incubation in Cellartis®medium, with CYP2C9 activity as the only exception. Glycogen storage and morphological integrity were maintained. Moreover, gene expression changes in hPCLS during incubation were limited and mostly related to cytoskeleton remodeling, fibrosis, and moderate oxidative stress. The expression of genes involved in drug transport, which is an important factor in determining the intracellular xenobiotic exposure, was also unchanged. Therefore, we conclude that hPCLS cultured in Cellartis®medium are a valuable human ex vivo model for toxicological and pharmacological studies that require prolonged xenobiotic exposure.
authors.affiliation: Division of Pharmacokinetics Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands. v.starokozhko@rug.nl.
authors.forename: Viktoriia
authors.identifier: http://orcid.org/0000-0002-7451-6793
authors.initials: V
authors.lastname: Starokozhko
authors.affiliation: Division of Pharmacokinetics Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands.
authors.forename: Suresh
authors.initials: S
authors.lastname: Vatakuti
authors.affiliation: Department of Clinical Pharmacy and Pharmacology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Bauke
authors.initials: B
authors.lastname: Schievink
authors.affiliation: Division of Pharmacokinetics Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands.
authors.forename: Marjolijn T
authors.initials: MT
authors.lastname: Merema
authors.affiliation: School of Biosciences, Systems Biology Research Center, University of Skövde, 54128, Skövde, Sweden.
authors.forename: Annika
authors.initials: A
authors.lastname: Asplund
authors.affiliation: School of Biosciences, Systems Biology Research Center, University of Skövde, 54128, Skövde, Sweden.
authors.forename: Jane
authors.initials: J
authors.lastname: Synnergren
authors.affiliation: Takara Bio Europe AB, Gothenburg, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Aspegren
authors.affiliation: Division of Pharmacokinetics Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands.
authors.forename: Geny M M
authors.initials: GMM
authors.lastname: Groothuis
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000418:Albumins / Q000096:biosynthesis; D002352:Carrier Proteins / Q000378:metabolism; D003470:Culture Media; D004798:Enzymes / Q000378:metabolism*; D005355:Fibrosis / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D008658:Inactivation, Metabolic; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D009924:Organ Culture Techniques / Q000379:methods*; D018384:Oxidative Stress / Q000235:genetics; D015262:Xenobiotics / Q000378:metabolism / Q000493:pharmacokinetics
chemical_list: D000418:Albumins; D002352:Carrier Proteins; D003470:Culture Media; D004798:Enzymes; D015262:Xenobiotics; D000255:Adenosine Triphosphate
keywords: Drug transport; Human precision-cut liver slices; Metabolism; Prolonged incubation; Transcriptomics