Cited by 160Drug-Induced Hepatotoxicity and ProtectionPharmacogenetics and Drug MetabolismAnimal testing and alternatives

Comparison of Drug Metabolism and Its Related Hepatotoxic Effects in HepaRG, Cryopreserved Human Hepatocytes, and HepG2 Cell Cultures

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Yokoyama, Yuichi, Sasaki, Yoshifumi, Terasaki, Natsuko, Kawataki, Taku, Takekawa, Koji, Iwase, Yumiko, Shimizu, Toshinobu, Sanoh, Seigo +1 more·2018·Biological and Pharmaceutical Bulletin

Abstract

Differentiated HepaRG cells maintain liver-specific functions such as drug-metabolizing enzymes. In this study, the feasibility of HepaRG cells as a human hepatocyte model for in vitro toxicity assessment was examined using selected hepatotoxic compounds. First, basal drug-metabolizing enzyme activities (CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4, uridine 5'-diphospho-glucuronosyltransferase [UGT], and sulfotransferases [SULT]) were measured in HepaRG, human hepatocytes, and HepG2 cells. Enzyme activities in differentiated HepaRG cells were comparable to those in human hepatocytes and much higher than those in HepG2 cells, except for SULT activity. Second, we examined the cytotoxicity of hepatotoxic compounds, acetaminophen (APAP), aflatoxin B1 (AFB1), cyclophosphamide (CPA), tamoxifen (TAM), and troglitazone (TGZ) in HepaRG cells and human hepatocytes. AFB1- and CPA-induced cytotoxicities against HepaRG cells were comparable to those against human hepatocytes. Furthermore, the cytotoxicities of these compounds were inhibited by 1-aminobenzotriazole (ABT), a broad CYP inhibitor, in both cells and were likely mediated by metabolic activation by CYP. Finally, toxicogenomics analysis of HepG2 and HepaRG cells after exposure to AFB1 and CPA revealed that numerous p53-related genes were upregulated- and the expression of these genes was greater in HepaRG than in HepG2 cells. These results suggest that gene expression profiles of HepaRG cells were affected more considerably by the toxic mechanisms of AFB1 and CPA than the profiles of HepG2 cells were. Therefore, our investigation shows that HepaRG cells could be useful human hepatic cellular models for toxicity studies.

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  • PHHs are the best in vitro hepatocyte models in current drug discovery research.

    “Primary (cryopreserved) human hepatocytes (PHHs) are the best in vitro hepatocyte models in current drug discovery research.7,8)”

    From Cryopreservation of Hepatocyte Organoid-Derived Cells Generated from Human iPS Cells (2025) · cites this paper as [8]

    The paper’s introductory passage (labelled Abstract in our copy; page unavailable) calls human hepatocytes the gold standard of human hepatic models for toxicological studies. It does not say cryopreserved hepatocytes are the best models across current drug discovery research: the Discussion notes their variability and says HepaRG cells may be more feasible for early drug-discovery screening. (LLM, conf=0.86)

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Research works printed but not linked: 4

  1. Westerink WM, Schoonen WG. Phase II enzyme levels in HepG2 cells and cryopreserved primary human hepatocytes and their in- duction in HepG2 cells. Toxicol. In Vitro, 21, 1592-1602 (2007). · DOI · Paper page
  2. Denissenko MF, Cahill J, Koudriakova TB, Gerber N, Pfeifer GP. Quantitation and mapping of aflatoxin B1-induced DNA damage in genomic DNA using aflatoxin B1-8,9-epoxide and microsomal acti- vation systems. Mutat. Res., 425, 205-211 (1999). · DOI
  3. McLean M, Dutton MF. Cellular interactions and metabolism of aflatoxin: an update. Pharmacol. Ther., 65, 163-192 (1995). · DOI
  4. Saito J, Okamura A, Takeuchi K, Hanioka K, Okada A, Ohata T. High content analysis assay for prediction of human hepatotoxicity in HepaRG and HepG2 cells. Toxicol. In Vitro, 33, 63-70 (2016). · DOI

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Authors
Yokoyama, Y., Sasaki, Y., Terasaki, N., Kawataki, T., Takekawa, K., Iwase, Y., Shimizu, T., Sanoh, S., & Ohta, S.
Affiliations
  • Hiroshima University (JP)
  • Tanabe Pharma Corporation (Japan) (JP)
Venue
Biological and Pharmaceutical Bulletin
Published
2018-05-01
Volume
41
Issue
5
Pages
722-732
ISSN
  • 0918-6158 (ISSN-L)
  • 1347-5215
OpenAlex
W2788132769
Cited by
160
References
37
Citations by year
2026: 12 · 2025: 19 · 2024: 23 · 2023: 16 · 2022: 17 · 2021: 23 · 2020: 37 · 2019: 8 · 2018: 5
FWCI
20.08
Type
journal article
Language
en
Open access
Locations
  • Biological and Pharmaceutical Bulletin (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
  • Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
Keywords
HepaRG cells, cryopreserved human hepatocytes, HepG2 cells, drug metabolism, hepatotoxicity, drug-metabolizing enzymes, CYP enzymes, UGT, sulfotransferase, aflatoxin B1, cyclophosphamide, paracetamol, tamoxifen, troglitazone, 1-aminobenzotriazole, cytochrome P450 inhibition, cytotoxicity, toxicogenomics, in vitro toxicity testing
MeSH terms
Cell Line, Cells, Cultured, Cytochrome P-450 Enzyme System, Drug Evaluation, Preclinical, Pharmaceutical Preparations, Humans, Glucuronosyltransferase, Sulfotransferases, Tumor Suppressor Protein p53, Toxicity Tests, Gene Expression Profiling, Hepatocytes, Hep G2 Cells, Drug-Related Side Effects and Adverse Reactions

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