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Cited by 37Liver Disease Diagnosis and TreatmentDiet, Metabolism, and DiseaseLipid metabolism and biosynthesis

A Comparison of Primary Human Hepatocytes and Hepatoma Cell Lines to Model the Effects of Fatty Acids, Fructose and Glucose on Liver Cell Lipid Accumulation

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Huggett, Zoë J., Smith, Alison, De Vivo, Nicola, Gomez, Dhanny, Jethwa, Preeti, Brameld, John M., Bennett, Andrew, Salter, Andrew M.·2022·Nutrients
DOI 10.3390/nu15010040PMC PMC9824391

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Abstract

Non-alcoholic fatty liver disease (NAFLD) begins with lipid accumulation within hepatocytes, but the relative contributions of different macronutrients is still unclear. We investigated the impact of fatty acids, glucose and fructose on lipid accumulation in primary human hepatocytes (PHH) and three different cell lines: HepG2 (human hepatoblastoma–derived cell line), Huh7 (human hepatocellular carcinoma cell line) and McA-RH7777 (McA, rat hepatocellular carcinoma cell line). Cells were treated for 48 h with fatty acids (0 or 200 μM), glucose (5 mM or 11 mM) and fructose (0 mM, 2 mM or 8 mM). Lipid accumulation was measured via Nile Red staining. All cell types accumulated lipid in response to fatty acids (p < 0.001). PHH and McA, but not HepG2 or Huh7 cells, accumulated more lipid with 11 mM glucose plus fatty acids (p = 0.004, fatty acid × glucose interaction, for both), but only PHH increased lipid accumulation in response to fructose (p < 0.001). Considerable variation was observed between PHH cells from different individuals. Lipid accumulation in PHH was increased by insulin (p = 0.003) with inter-individual variability. Similarly, insulin increased lipid accumulation in both HepG2 and McA cells, with a bigger response in McA in the presence of fatty acids (p < 0.001 for fatty acid × insulin). McA were more insulin sensitive than either HepG2 or Huh7 cells in terms of AKT phosphorylation (p < 0.001 insulin × cell type interaction). Hence, glucose and fructose can contribute to the accumulation of lipid in PHH with considerable inter-individual variation, but hepatoma cell lines are not good models of PHH.

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  • HepG2 expresses high-affinity hexokinase II (HK2) instead of glucokinase (GCK), maintaining high glycolysis even at low glucose.

    “While mature hepatocytes express glucokinase (GCK, hexokinase IV), whose activity is regulated by physiological glucose concentrations and insulin, GCK expression is suppressed in HepG2 cells, which produce high-affinity hexokinase II (HK2) [116,117,118].”

    From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [116]

    The Discussion reports that HepG2 cells express hexokinase II rather than the glucokinase expressed in primary hepatocytes. It does not describe hexokinase II as high-affinity or establish that this substitution maintains high glycolysis at low glucose. (LLM, conf=0.86)

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Authors
Huggett, Z. J., Smith, A., De Vivo, N., Gomez, D., Jethwa, P., Brameld, J. M., Bennett, A., & Salter, A. M.
Affiliations
  • University of Nottingham (GB)
  • Fund for the Replacement of Animals in Medical Experiments (FRAME) Laboratory, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, Nottinghamshire, UK
  • Division of Food, Nutrition and Dietetics, School of Biosciences, Sutton Bonington Campus, University of Nottingham, Loughborough LE12 5RD, Leicestershire, UK
ORCID
Venue
Nutrients
Publisher
MDPI AG
Published
2022-12-21
Volume
15
Issue
1
Pages
40
ISSN
2072-6643 (ISSN-L)
DOI
10.3390/nu15010040
PMC
PMC9824391
OpenAlex
W4312172058
Cited by
37
References
37
Citations by year
2026: 11 · 2025: 15 · 2024: 10 · 2023: 1
FWCI
4.69
Type
journal article
Language
en
Open access
Licence
Locations
  • Nutrients (publishedVersion, cc-by)
  • Preprints.org (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Repository@Nottingham (University of Nottingham) (publishedVersion, cc-by)
  • Repository@Nottingham (University of Nottingham) (submittedVersion, cc-by)
Funders
Rank Prize Fund
Topics
  • Health Sciences › Medicine › Epidemiology › Liver Disease Diagnosis and Treatment
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Diet, Metabolism, and Disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
Keywords
non-alcoholic fatty liver disease, hepatic lipid accumulation, primary human hepatocytes, hepatoma cell lines, fatty acids, fructose, glucose, insulin sensitivity, Akt phosphorylation, interindividual variability, HepG2 cells, Huh7 cells, cell line models
MeSH terms
Animals, Cell Line, Fatty Acids, Fructose, Glucose, Carcinoma, Hepatocellular, Humans, Liver Neoplasms, Hepatocytes, Lipid Metabolism, Rats, Hep G2 Cells, Insulins, Non-alcoholic Fatty Liver Disease

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