title: Novel human hepatic organoid model enables testing of drug-induced liver fibrosis in vitro.
abstract: Current models for in vitro fibrosis consist of simple mono-layer cultures of rodent hepatic stellate cells (HSC), ignoring the role of hepatocyte injury. We aimed to develop a method allowing the detection of hepatocyte-mediated and drug-induced liver fibrosis. We used HepaRG (Hep) and primary human HSCs cultured as 3D spheroids in 96-well plates. These resulting scaffold-free organoids were characterized for CYP induction, albumin secretion, and hepatocyte and HSC-specific gene expression by qPCR. The metabolic competence of the organoid over 21 days allows activation of HSCs in the organoid in a drug- and hepatocyte-dependent manner. After a single dose or repeated exposure for 14 days to the pro-fibrotic compounds Allyl alcohol and Methotrexate, hepatic organoids display fibrotic features such as HSC activation, collagen secretion and deposition. Acetaminophen was identified by these organoids as an inducer of hepatotoxic-mediated HSC activation which was confirmed in vivo in mice. This novel hepatic organoid culture model is the first that can detect hepatocyte-dependent and compound-induced HSC activation, thereby representing an important step forward towards in vitro compound testing for drug-induced liver fibrosis.
authors.affiliation: Liver Cell Biology Laboratory, Vrije Universiteit Brussel (VUB), Belgium. Electronic address: sofia.batista.leite@vub.ac.be.
authors.forename: Sofia B
authors.initials: SB
authors.lastname: Leite
authors.affiliation: Liver Cell Biology Laboratory, Vrije Universiteit Brussel (VUB), Belgium.
authors.forename: Tiffany
authors.initials: T
authors.lastname: Roosens
authors.affiliation: Liver Cell Biology Laboratory, Vrije Universiteit Brussel (VUB), Belgium.
authors.forename: Adil
authors.initials: A
authors.lastname: El Taghdouini
authors.affiliation: Liver Cell Biology Laboratory, Vrije Universiteit Brussel (VUB), Belgium.
authors.forename: Inge
authors.initials: I
authors.lastname: Mannaerts
authors.affiliation: Liver Cell Biology Laboratory, Vrije Universiteit Brussel (VUB), Belgium.
authors.forename: Ayla J
authors.initials: AJ
authors.lastname: Smout
authors.affiliation: Laboratory of Pediatric Hepatology and Cell Therapy, Institute of Experimental and Clinical Research (IREC), Université Catholique de Louvain (UCL), Belgium.
authors.forename: Mustapha
authors.initials: M
authors.lastname: Najimi
authors.affiliation: Laboratory of Pediatric Hepatology and Cell Therapy, Institute of Experimental and Clinical Research (IREC), Université Catholique de Louvain (UCL), Belgium.
authors.forename: Etienne
authors.initials: E
authors.lastname: Sokal
authors.affiliation: Biochemical Engineering Institute, Saarland University, Germany.
authors.forename: Fozia
authors.initials: F
authors.lastname: Noor
authors.affiliation: Biopredic International, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Chesne
authors.affiliation: Liver Cell Biology Laboratory, Vrije Universiteit Brussel (VUB), Belgium. Electronic address: lvgrunsv@vub.ac.be.
authors.forename: Leo A
authors.initials: LA
authors.lastname: van Grunsven
mesh_terms: D002460:Cell Line; D006801:Humans; D008099:Liver / Q000473:pathology*; D008103:Liver Cirrhosis / Q000139:chemically induced*; D008954:Models, Biological
keywords: APAP; Allyl alcohol; DILI; HepaRG; Hepatic stellate cell; Hepatocyte; In vitro; Liver fibrosis; Methotrexate; Organoid