title: Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes.
abstract: OBJECTIVE
  Hepatocyte-like cells (HLCs), differentiated from pluripotent stem cells by the use of soluble factors, can model human liver function and toxicity. However, at present HLC maturity and whether any deficit represents a true fetal state or aberrant differentiation is unclear and compounded by comparison to potentially deteriorated adult hepatocytes. Therefore, we generated HLCs from multiple lineages, using two different protocols, for direct comparison with fresh fetal and adult hepatocytes.
  
  
  METHODS
  Protocols were developed for robust differentiation. Multiple transcript, protein and functional analyses compared HLCs to fresh human fetal and adult hepatocytes.
  
  
  RESULTS
  HLCs were comparable to those of other laboratories by multiple parameters. Transcriptional changes during differentiation mimicked human embryogenesis and showed more similarity to pericentral than periportal hepatocytes. Unbiased proteomics demonstrated greater proximity to liver than 30 other human organs or tissues. However, by comparison to fresh material, HLC maturity was proven by transcript, protein and function to be fetal-like and short of the adult phenotype. The expression of 81% phase 1 enzymes in HLCs was significantly upregulated and half were statistically not different from fetal hepatocytes. HLCs secreted albumin and metabolized testosterone (CYP3A) and dextrorphan (CYP2D6) like fetal hepatocytes. In seven bespoke tests, devised by principal components analysis to distinguish fetal from adult hepatocytes, HLCs from two different source laboratories consistently demonstrated fetal characteristics.
  
  
  CONCLUSIONS
  HLCs from different sources are broadly comparable with unbiased proteomic evidence for faithful differentiation down the liver lineage. This current phenotype mimics human fetal rather than adult hepatocytes.
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Melissa
authors.initials: M
authors.lastname: Baxter
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Withey
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Sean
authors.initials: S
authors.lastname: Harrison
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, Anne McLaren Institute for Regenerative Medicine, Department of Surgery, Robinson Way, Cambridge CB2 0SZ, UK; Wellcome Trust Sanger Institute, Hinxton CB10 1SA, UK.
authors.forename: Charis-Patricia
authors.initials: CP
authors.lastname: Segeritz
authors.affiliation: Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Fang
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Atkinson-Dell
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK; Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Cliff
authors.initials: C
authors.lastname: Rowe
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK; Bioinformatics, Faculty of Life Sciences, Michael Smith Building, Oxford Road, Manchester, UK.
authors.forename: Dave T
authors.initials: DT
authors.lastname: Gerrard
authors.affiliation: Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Rowena
authors.initials: R
authors.lastname: Sison-Young
authors.affiliation: Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Roz
authors.initials: R
authors.lastname: Jenkins
authors.affiliation: Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Joanne
authors.initials: J
authors.lastname: Henry
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Berry
authors.affiliation: Stem Cell Research Group, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Lisa
authors.initials: L
authors.lastname: Mohamet
authors.affiliation: Human Genetics Division, University of Southampton, Southampton General Hospital, Tremona Road, Southampton, UK.
authors.forename: Marie
authors.initials: M
authors.lastname: Best
authors.affiliation: North Western Hepatobiliary Unit, Aintree University Hospital NHS Foundation Trust, Longmoor Lane, Liverpool L9 7AL, UK.
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Fenwick
authors.affiliation: North Western Hepatobiliary Unit, Aintree University Hospital NHS Foundation Trust, Longmoor Lane, Liverpool L9 7AL, UK.
authors.forename: Hassan
authors.initials: H
authors.lastname: Malik
authors.affiliation: Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Neil R
authors.initials: NR
authors.lastname: Kitteringham
authors.affiliation: Department of Pharmacology & Therapeutics and MRC Centre for Drug Safety Science, University of Liverpool, Sherrington Building, Ashton Street, Liverpool, UK.
authors.forename: Chris E
authors.initials: CE
authors.lastname: Goldring
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK.
authors.forename: Karen
authors.initials: K
authors.lastname: Piper Hanley
authors.affiliation: Wellcome Trust-Medical Research Council Stem Cell Institute, Anne McLaren Institute for Regenerative Medicine, Department of Surgery, Robinson Way, Cambridge CB2 0SZ, UK; Wellcome Trust Sanger Institute, Hinxton CB10 1SA, UK.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Vallier
authors.affiliation: Centre for Endocrinology & Diabetes, Institute of Human Development, Faculty of Medical & Human Sciences, University of Manchester, Manchester Academic Health Science Centre, AV Hill Building, Oxford Road, Manchester, UK; Endocrinology Department, Central Manchester University Hospitals NHS Foundation Trust, Grafton St, Manchester, UK. Electronic address: Neil.Hanley@manchester.ac.uk.
authors.forename: Neil A
authors.initials: NA
authors.lastname: Hanley
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D053686:Fetal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D055442:Metabolome; D008954:Models, Biological; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism
chemical_list: D020543:Proteome
keywords: Embryo; Hepatic; Hepatotoxicity; Human embryonic stem cell; Liver