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Cited by 21Liver Disease Diagnosis and TreatmentMitochondrial Function and PathologyAutophagy in Disease and Therapy

Comparison of HepaRG and HepG2 cell lines to model mitochondrial respiratory adaptations in non‑alcoholic fatty liver disease

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Maseko, Tumisang, Elkalaf, Moustafa, Peterová, Eva, Lotková, Halka, Staňková, Pavla, Melek, Jan, Dušek, Jan, Žádníková, Petra +5 more·2023·International Journal of Molecular Medicine
DOI 10.3892/ijmm.2023.5342PMC PMC10781417
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Abstract

Although some clinical studies have reported increased mitochondrial respiration in patients with fatty liver and early non‑alcoholic steatohepatitis (NASH), there is a lack of in vitro models of non‑alcoholic fatty liver disease (NAFLD) with similar findings. Despite being the most commonly used immortalized cell line for in vitro models of NAFLD, HepG2 cells exposed to free fatty acids (FFAs) exhibit a decreased mitochondrial respiration. On the other hand, the use of HepaRG cells to study mitochondrial respiratory changes following exposure to FFAs has not yet been fully explored. Therefore, the present study aimed to assess cellular energy metabolism, particularly mitochondrial respiration, and lipotoxicity in FFA‑treated HepaRG and HepG2 cells. HepaRG and HepG2 cells were exposed to FFAs, followed by comparative analyses that examained cellular metabolism, mitochondrial respiratory enzyme activities, mitochondrial morphology, lipotoxicity, the mRNA expression of selected genes and triacylglycerol (TAG) accumulation. FFAs stimulated mitochondrial respiration and glycolysis in HepaRG cells, but not in HepG2 cells. Stimulated complex I, II‑driven respiration and β‑oxidation were linked to increased complex I and II activities in FFA‑treated HepaRG cells, but not in FFA‑treated HepG2 cells. Exposure to FFAs disrupted mitochondrial morphology in both HepaRG and HepG2 cells. Lipotoxicity was induced to a greater extent in FFA‑treated HepaRG cells than in FFA‑treated HepG2 cells. TAG accumulation was less prominent in HepaRG cells than in HepG2 cells. On the whole, the present study demonstrates that stimulated mitochondrial respiration is associated with lipotoxicity in FFA‑treated HepaRG cells, but not in FFA‑treated HepG2 cells. These findings suggest that HepaRG cells are more suitable for assessing mitochondrial respiratory adaptations in the developed in vitro model of early‑stage NASH.

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  • For tasks related to disease progression (inflammation, fibrosis), as well as for rigorous studies of drug metabolism and toxicity, HepG2 should be used with caution, preferably as part of more complex systems (co-cultures, 3D platforms) or in combination with alternative models (HepaRG, primary hepatocytes).

    “For tasks related to disease progression (inflammation, fibrosis), as well as for rigorous studies of drug metabolism and toxicity, HepG2 should be used with caution, preferably as part of more complex systems (co-cultures, 3D platforms) or in combination with alternative models (HepaRG, primary hepatocytes) [158,159,160,161].”

    From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [158]

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Authors
Maseko, T., Elkalaf, M., Peterová, E., Lotková, H., Staňková, P., Melek, J., Dušek, J., Žádníková, P., Čížková, D., Bezrouk, A., Pávek, P., Červinková, Z., & Kučera, O.
Affiliations
Charles University (CZ)
Corresponding author
Kučera, Otto
ORCID
Venue
International Journal of Molecular Medicine
Publisher
Spandidos Publications
Published
2023-12-22
Volume
53
Issue
2
ISSN
  • 1107-3756 (ISSN-L)
  • 1791-244X
DOI
10.3892/ijmm.2023.5342
PMC
PMC10781417
OpenAlex
W4390105489
Cited by
21
References
5
Citations by year
2026: 11 · 2025: 5 · 2024: 5
FWCI
4.19
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • International Journal of Molecular Medicine (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Univerzita Karlova v Praze (/0010046, SVV-2023-260656)
  • Grantová Agentura, Univerzita Karlova
  • European Commission (CZ.02.1.01/0.0/0.0/18_069/0010046, CZ.02.1.01)
Topics
  • Health Sciences › Medicine › Epidemiology › Liver Disease Diagnosis and Treatment
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
Keywords
HepaRG cells, HepG2 cells, non-alcoholic fatty liver disease, nonalcoholic steatohepatitis, mitochondrial respiration, free fatty acids, lipotoxicity, mitochondrial morphology, triglyceride accumulation, glycolysis, fatty acid beta-oxidation, cellular energy metabolism, in vitro disease model
MeSH terms
Cell Line, Fatty Acids, Nonesterified, Humans, Mitochondria, Respiration, Triglycerides, Hep G2 Cells, Non-alcoholic Fatty Liver Disease

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