title: Small-Molecule-Directed Hepatocyte-Like Cell Differentiation of Human Pluripotent Stem Cells.
abstract: Hepatocyte-like cells (HLCs) generated in vitro from human pluripotent stem cells (hPSCs) provide an invaluable resource for basic research, regenerative medicine, drug screening, toxicology, and modeling of liver disease and development. This unit describes a small-molecule-driven protocol for in vitro differentiation of hPSCs into HLCs without the use of growth factors. hPSCs are coaxed through a developmentally relevant route via the primitive streak to definitive endoderm (DE) using the small molecule CHIR99021 (a Wnt agonist), replacing the conventional growth factors Wnt3A and activin A. The small-molecule-derived DE is then differentiated to hepatoblast-like cells in the presence of dimethyl sulfoxide. The resulting hepatoblasts are then differentiated to HLCs with N-hexanoic-Tyr, Ile-6 aminohexanoic amide (Dihexa, a hepatocyte growth factor agonist) and dexamethasone. The protocol provides an efficient and reproducible procedure for differentiation of hPSCs into HLCs utilizing small molecules. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Norway.|Norwegian Center for Stem Cell Research, Oslo, Norway.
authors.forename: Santosh
authors.initials: S
authors.lastname: Mathapati
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Norway.|Norwegian Center for Stem Cell Research, Oslo, Norway.
authors.forename: Richard
authors.initials: R
authors.lastname: Siller
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Norway.|Department of Medical Genetics, Oslo University Hospital, Ullevål, Oslo, Norway.
authors.forename: Agata A R
authors.initials: AA
authors.lastname: Impellizzeri
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Norway.
authors.forename: Max
authors.initials: M
authors.lastname: Lycke
authors.affiliation: Department of Pediatric Research, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, Norway.
authors.forename: Karianne
authors.initials: K
authors.lastname: Vegheim
authors.affiliation: Department of Pediatric Research, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, Norway.
authors.forename: Runar
authors.initials: R
authors.lastname: Almaas
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Norway.|Norwegian Center for Stem Cell Research, Oslo, Norway.|Institute of Immunology, Oslo University Hospital, Oslo, Norway.
authors.forename: Gareth J
authors.initials: GJ
authors.lastname: Sullivan
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects*; D004707:Endoderm / Q000166:cytology; D061252:Feeder Cells / Q000166:cytology / Q000187:drug effects; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology*; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D054852:Small Molecule Libraries
keywords: definitive endoderm; hESCs; hepatocyte; hiPSCs; small molecules