title: Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease.
abstract: Liver biology and function, drug-induced liver injury (DILI) and liver diseases are difficult to study using current in vitro models such as primary human hepatocyte (PHH) monolayer cultures, as their rapid de-differentiation restricts their usefulness substantially. Thus, we have developed and extensively characterized an easily scalable 3D PHH spheroid system in chemically-defined, serum-free conditions. Using whole proteome analyses, we found that PHH spheroids cultured this way were similar to the liver in vivo and even retained their inter-individual variability. Furthermore, PHH spheroids remained phenotypically stable and retained morphology, viability, and hepatocyte-specific functions for culture periods of at least 5 weeks. We show that under chronic exposure, the sensitivity of the hepatocytes drastically increased and toxicity of a set of hepatotoxins was detected at clinically relevant concentrations. An interesting example was the chronic toxicity of fialuridine for which hepatotoxicity was mimicked after repeated-dosing in the PHH spheroid model, not possible to detect using previous in vitro systems. Additionally, we provide proof-of-principle that PHH spheroids can reflect liver pathologies such as cholestasis, steatosis and viral hepatitis. Combined, our results demonstrate that the PHH spheroid system presented here constitutes a versatile and promising in vitro system to study liver function, liver diseases, drug targets and long-term DILI.
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Catherine C
authors.initials: CC
authors.lastname: Bell
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Delilah F G
authors.initials: DF
authors.lastname: Hendriks
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Sabrina M L
authors.initials: SM
authors.lastname: Moro
authors.affiliation: Department of Clinical Science, Intervention and Technology, Karolinska University Hospital Huddinge, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Ewa
authors.initials: E
authors.lastname: Ellis
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, UK.
authors.forename: Joanne
authors.initials: J
authors.lastname: Walsh
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Renblom
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Lisa
authors.initials: L
authors.lastname: Fredriksson Puigvert
authors.affiliation: Janssen Pharmaceutical Companies of Johnson &Johnson, Department of Pharmacokinetics, Dynamics and Metabolism, Beerse, Belgium.
authors.forename: Anita C A
authors.initials: AC
authors.lastname: Dankers
authors.affiliation: Janssen Pharmaceutical Companies of Johnson &Johnson, Department of Pharmacokinetics, Dynamics and Metabolism, Beerse, Belgium.
authors.forename: Frank
authors.initials: F
authors.lastname: Jacobs
authors.affiliation: Janssen Pharmaceutical Companies of Johnson &Johnson, Department of Pharmacokinetics, Dynamics and Metabolism, Beerse, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Snoeys
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, UK.
authors.forename: Rowena L
authors.initials: RL
authors.lastname: Sison-Young
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, UK.
authors.forename: Rosalind E
authors.initials: RE
authors.lastname: Jenkins
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Åsa
authors.initials: Å
authors.lastname: Nordling
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Souren
authors.initials: S
authors.lastname: Mkrtchian
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, UK.
authors.forename: B Kevin
authors.initials: BK
authors.lastname: Park
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, UK.
authors.forename: Neil R
authors.initials: NR
authors.lastname: Kitteringham
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, UK.
authors.forename: Christopher E P
authors.initials: CE
authors.lastname: Goldring
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Volker M
authors.initials: VM
authors.lastname: Lauschke
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Magnus
authors.initials: M
authors.lastname: Ingelman-Sundberg
mesh_terms: D001086:Arabinofuranosyluracil / Q000031:analogs & derivatives / Q000633:toxicity; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology* / Q000503:physiopathology*; D022781:Hepatocytes / Q000187:drug effects* / Q000502:physiology*; D006801:Humans; D008954:Models, Biological; D000075082:Proof of Concept Study; D020543:Proteome / Q000032:analysis; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000502:physiology*
chemical_list: D020543:Proteome; D001086:Arabinofuranosyluracil; C043457:fialuridine