title: Characterization of lipid metabolism in a novel immortalized human hepatocyte cell line.
abstract: The development of hepatocyte cell models that represent fatty acid partitioning within the human liver would be beneficial for the study of the development and progression of nonalcoholic fatty liver disease (NAFLD). We sought to develop and characterize a novel human liver cell line (LIV0APOLY) to establish a model of lipid accumulation using a physiological mixture of fatty acids under low- and high-glucose conditions. LIV0APOLY cells were compared with a well-established cell line (HepG2) and, where possible, primary human hepatocytes. LIV0APOLY cells were found to proliferate and express some mature liver markers and were wild type for the PNPLA3 (rs738409) gene, whereas HepG2 cells carried the Ile(148)Met variant that is positively associated with liver fat content. Intracellular triglyceride content was higher in HepG2 than in LIV0APOLY cells; exposure to high glucose and/or exogenous fatty acids increased intracellular triglyceride in both cell lines. Triglyceride concentrations in media were higher from LIV0APOLY compared with HepG2 cells. Culturing LIV0APOLY cells in high glucose increased a marker of endoplasmic reticulum stress and attenuated insulin-stimulated Akt phosphorylation whereas low glucose and exogenous fatty acids increased AMPK phosphorylation. Although LIV0APOLY cells and primary hepatocytes stored similar amounts of exogenous fatty acids as triglyceride, more exogenous fatty acids were partitioned toward oxidation in the LIV0APOLY cells than in primary hepatocytes. LIV0APOLY cells offer the potential to be a renewable cellular model for studying the effects of exogenous metabolic substrates on fatty acid partitioning; however, their usefulness as a model of lipoprotein metabolism needs to be further explored.
authors.affiliation: Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, United Kingdom;
authors.forename: Charlotte J
authors.initials: CJ
authors.lastname: Green
authors.affiliation: ReNeuron Group, Guildford, Surrey, United Kingdom;
authors.forename: Deborah
authors.initials: D
authors.lastname: Johnson
authors.affiliation: ReNeuron Group, Guildford, Surrey, United Kingdom;
authors.forename: Harsh D
authors.initials: HD
authors.lastname: Amin
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, The Women's Centre, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom;
authors.forename: Pamela
authors.initials: P
authors.lastname: Sivathondan
authors.affiliation: Department of Hepatobiliary and Pancreatic Surgery, Oxford University Hospital NHS Trust, Churchill Hospital, Oxford, United Kingdom; and.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Silva
authors.affiliation: Department of Cellular Pathology, Oxford University Hospitals, Oxford, United Kingdom.
authors.forename: Lai Mun
authors.initials: LM
authors.lastname: Wang
authors.affiliation: ReNeuron Group, Guildford, Surrey, United Kingdom;
authors.forename: Lara
authors.initials: L
authors.lastname: Stevanato
authors.affiliation: Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, United Kingdom;
authors.forename: Catriona A
authors.initials: CA
authors.lastname: McNeil
authors.affiliation: ReNeuron Group, Guildford, Surrey, United Kingdom;
authors.forename: Erik A
authors.initials: EA
authors.lastname: Miljan
authors.affiliation: ReNeuron Group, Guildford, Surrey, United Kingdom;
authors.forename: John D
authors.initials: JD
authors.lastname: Sinden
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, The Women's Centre, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom;
authors.forename: Karl J
authors.initials: KJ
authors.lastname: Morten
authors.affiliation: Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, United Kingdom; leanne.hodson@ocdem.ox.ac.uk.
authors.forename: Leanne
authors.identifier: http://orcid.org/0000-0002-2648-6526
authors.initials: L
authors.lastname: Hodson
mesh_terms: D002460:Cell Line; D059865:Endoplasmic Reticulum Stress; D005227:Fatty Acids / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D008049:Lipase / Q000235:genetics; D050356:Lipid Metabolism; D008099:Liver / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics; D065626:Non-alcoholic Fatty Liver Disease / Q000378:metabolism*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism*; D000217:Acyltransferases; D054512:Phospholipases A2, Calcium-Independent
chemical_list: D005227:Fatty Acids; D005947:Glucose; D007328:Insulin; D008049:Lipase; D008565:Membrane Proteins; D051057:Proto-Oncogene Proteins c-akt; D014280:Triglycerides; C533682:PNPLA3 protein, human; D000217:Acyltransferases; D054512:Phospholipases A2, Calcium-Independent
keywords: NAFLD; PNPLA3; hepatocyte; human; triglyceride