Nutrient environment improves drug metabolic activity in human iPSC-derived hepatocytes and HepG2
Open accessAbstract
Induced pluripotent stem cells (iPSCs) have emerged as a transformative tool in regenerative medicine, in liver research. The perspective of a stable and functional source of hepatocytes has led to developing protocols for human iPSC-derived hepatocytes-like cells (HLCs). Yet, hepatic models remain one of most challenging systems to functionally reproduce with iPSCs, due to its resulting limited metabolic function. Using an adapted nutrient regimen, two human hepatocyte models were characterized: HLCs (derived from iPSCs) and metabolically active HepG2 (mHepG2, derived from the cell line HepG2), for their drug metabolism activity. In these cell systems, the transcriptome, proteome, and metabolome of 11 drug-relevant cytochrome P450 (CYP) isoenzymes were studied. A liquid chromatography–mass spectrometry (LC–MS)-based metabolomics approach, using model drugs as isoenzyme reporters, was applied, achieving a comprehensive overview of mHepG2 and HLCs drug metabolism. Drugs used in this study to characterize xenobiotic machinery were: bupropion (25 µM), phenacetin (30 µM), rosiglitazone (10 µM), diclofenac (75 µM), dextromethorphan (15 µM), chlorzoxazone (60 µM), midazolam (15 µM), benzydamine (15 µM), coumarin (250 µM) and 7-ethoxycoumarin (60 µM). Being HepG2 notorious for its limited metabolic capacity, our study raises mHepG2 as a highly performant cell model, with activity on 8 drug-metabolizing CYPs. Modulation by nutrient environment in improving metabolic function of in vitro models is here proven as a key determinant. Likewise, HLCs hold the widest CYP coverage at the transcript level and were able to cope with a wide variety of chemical insults, making them a promising model for personalized metabolic studies.
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Under amino acid-enriched conditions, a metabolically active HepG2 (mHepG2) variant has been described that exhibits activity of eight drug-metabolizing CYPs (CYP1A2, 2A6, 2B6, 2C8, 2C9, 2D6, 2E1, and 3A4), as confirmed at the transcriptional, proteomic, and metabolic levels; however, this system requires a lengthy preparation period (22 days of cultivation) and does not eliminate the line's tumor
“Under amino acid-enriched conditions, a metabolically active HepG2 (mHepG2) variant has been described that exhibits activity of eight drug-metabolizing CYPs (CYP1A2, 2A6, 2B6, 2C8, 2C9, 2D6, 2E1, and 3A4), as confirmed at the transcriptional, proteomic, and metabolic levels. However, this system requires a lengthy preparation period (22 days of cultivation) and does not eliminate the line’s tumor”
From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [168]
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- Authors
- Pozo Garcia, V., Çobanoğlu, T. S., Hammer, H. S., Carlota, R., Holm, K., Verfaillie, C., Poetz, O., Jennings, P., & Moco, S.
- Affiliations
- Vrije Universiteit Amsterdam (NL)
- Signatope GmbH (Germany) (DE)
- Leuven Stem Cell Institute (BE)
- Natural and Medical Sciences Institute (DE)
- Corresponding author
- Moco, Sofia
- ORCID
- Venue
- Archives of Toxicology
- Published
- 2025-08-12
- Volume
- 99
- Issue
- 11
- Pages
- 4493-4511
- ISSN
- 0340-5761 (ISSN-L)
- 1432-0738
- PMC
- PMC12477091
- OpenAlex
- W4413109370
- Cited by
- 15
- References
- 68
- Citations by year
- 2026: 13 · 2025: 2
- FWCI
- 14.55
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Archives of Toxicology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by)
- VU Research Portal (publishedVersion, cc-by)
- Funders
- EU RISK-HUNT3R (964537)
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- iPSC-derived hepatocytes, HepG2 cells, drug metabolism, cytochrome P450, nutrient environment, hepatocyte-like cells, metabolomics, xenobiotic metabolism
- MeSH terms
- Nutrients, Chromatography, Liquid, Cytochrome P-450 Enzyme System, Humans, Xenobiotics, Hepatocytes, Metabolomics, Metabolome, Hep G2 Cells, Induced Pluripotent Stem Cells, Transcriptome
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