title: Comparison of Liver Cell Models Using the Basel Phenotyping Cocktail.
abstract: Currently used hepatocyte cell systems forin vitroassessment of drug metabolism include hepatoma cell lines and primary human hepatocyte (PHH) cultures. We investigated the suitability of the validatedin vivoBasel phenotyping cocktail (caffeine [CYP1A2], efavirenz [CYP2B6], losartan [CYP2C9], omeprazole [CYP2C19], metoprolol [CYP2D6], midazolam [CYP3A4])in vitroand characterized four hepatocyte cell systems (HepG2 cells, HepaRG cells, and primary cryopreserved human hepatocytes in 2-dimensional [2D] culture or in 3D-spheroid co-culture) regarding basal metabolism and CYP inducibility. Under non-induced conditions, all CYP activities could be determined in 3D-PHH, CYP2B6, CYP2C19, CYP2D6, and CYP3A4 in 2D-PHH and HepaRG, and CYP2C19 and CYP3A4 in HepG2 cells. The highest non-induced CYP activities were observed in 3D-PHH and HepaRG cells. mRNA expression was at least four-fold higher for all CYPs in 3D-PHH compared to the other cell systems. After treatment with 20 μM rifampicin, mRNA increased 3- to 50-fold for all CYPs except CYP1A2 and 2D6 for HepaRG and 3D-PHH, 4-fold (CYP2B6) and 17-fold (CYP3A4) for 2D-PHH and four-fold (CYP3A4) for HepG2. In 3D-PHH at least a two-fold increase in CYP activity was observed for all inducible CYP isoforms while CYP1A2 and CYP2C9 activity did not increase in 2D-PHH and HepaRG. CYP inducibility assessedin vivousing the same phenotyping probes was also best reflected by the 3D-PHH model. Our studies show that 3D-PHH and (with some limitations) HepaRG are suitable cell systems for assessing drug metabolism and CYP inductionin vitro. HepG2 cells are less suited to assess CYP induction of the 2C and 3A family. The Basel phenotyping cocktail is suitable for the assessment of CYP activity and induction alsoin vitro.
authors.affiliation: Division of Clinical Pharmacology and Toxicology, University Hospital BaselBasel, Switzerland; Department of Biomedicine, University of BaselBasel, Switzerland.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Berger
authors.affiliation: Division of Clinical Pharmacology and Toxicology, University Hospital BaselBasel, Switzerland; Department of Biomedicine, University of BaselBasel, Switzerland.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Donzelli
authors.affiliation: Division of Clinical Pharmacology and Toxicology, University Hospital BaselBasel, Switzerland; Department of Biomedicine, University of BaselBasel, Switzerland.
authors.forename: Swarna
authors.initials: S
authors.lastname: Maseneni
authors.affiliation: Roche Innovation Center Basel, Pharmaceutical Sciences, Hoffmann-La Roche Ltd. Basel, Switzerland.
authors.forename: Franziska
authors.initials: F
authors.lastname: Boess
authors.affiliation: Roche Innovation Center Basel, Pharmaceutical Sciences, Hoffmann-La Roche Ltd. Basel, Switzerland.
authors.forename: Adrian
authors.initials: A
authors.lastname: Roth
authors.affiliation: Division of Clinical Pharmacology and Toxicology, University Hospital BaselBasel, Switzerland; Department of Biomedicine, University of BaselBasel, Switzerland; Swiss Center for Applied Human ToxicologyBasel, Switzerland.
authors.forename: Stephan
authors.initials: S
authors.lastname: Krähenbühl
authors.affiliation: Division of Clinical Pharmacology and Toxicology, University Hospital BaselBasel, Switzerland; Department of Biomedicine, University of BaselBasel, Switzerland.
authors.forename: Manuel
authors.initials: M
authors.lastname: Haschke
keywords: 3-dimensional spheroid primary human hepatocyte culture; CYP induction; basel cocktail; cytochrome P450 (CYP); liquid chromatography tandem mass spectrometry