Co-culture with mouse embryonic fibroblasts improves maintenance of metabolic function of human small hepatocyte progenitor cells
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has been described in rodents and recently in humans. These cells are capable of engrafting in injured livers, however, they display de-differentiated morphology and reduced xenobiotic metabolism activity in culture over passages. Here we report that SHPCs derived from adult primary human hepatocytes (PHHs) and cultured on mouse embryonic fibroblasts (MEFs) not only display differentiated morphology and exhibit gene expression profiles similar to adult PHHs, but importantly, they retain their phenotype over several passages. Further, unlike previous reports, where extensive manipulations of culture conditions are required to convert SHPCs to metabolically functional hepatocytes, SHPCs in our co-culture system maintain expression of xenobiotic metabolism-associated genes. We show that SHPCs in co-culture are able to perform xenobiotic metabolism at rates equal to their parent PHHs as evidenced by the metabolism of acetaminophen to all of its major metabolites. In summary, we present an improved co-culture system that allows generation of SHPCs from adult PHHs that maintain their differentiated phenotype over multiple passages. Our findings would be useful for expansion of limited PHHs for use in studies of drug metabolism and toxicity testing.
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Under optimized conditions, PHH-derived SHPCs exhibited polarized morphology, functional bile canaliculi, persistent expression of hepatocyte-specific transcription factors and transporters, and maintained CYP450 activity, leading to drug metabolism capacity comparable to that of primary hepatocytes.
“Under optimized conditions, PHH-derived SHPCs exhibited polarized morphology, functional bile canaliculi, persistent expression of hepatocyte-specific transcription factors, and transporters and maintained CYP450 activity, leading to drug metabolism capacity comparable to that of PHs.”
From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [32]
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Experimental — accuracy not yet measuredSmall hepatocyte progenitor-like cells were derived from adult primary human hepatocytes by direct co-culture with irradiated mouse embryonic fibroblasts, which provided feeder-dependent signals that triggered proliferation while maintaining necessary hepatocyte features.
“In this regard, small hepatocyte progenitor-like cells (SHPCs) were derived from adult PHHs by direct co-culture with irradiated mouse embryonic fibroblasts (MEFs), which provided feeder-dependent signals that triggered proliferation while maintaining necessary hepatocyte features [32].”
From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [32]
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- Authors
- Sengupta, S., Johnson, B., Seirup, M., Ardalani, H., Duffin, B., Barrett-Wilt, G. A., Stewart, R., & Thomson, J. A.
- Affiliations
- Morgridge Institute for Research (US)
- University of Wisconsin–Madison (US)
- Corresponding author
- Thomson, James A.
- Venue
- Current Research in Toxicology
- Publisher
- Elsevier BV
- Published
- 2020-06
- Volume
- 1
- Pages
- 70-84
- ISSN
- 2666-027X (ISSN-L)
- PMC
- PMC8320630
- OpenAlex
- W3080092126
- Cited by
- 9
- References
- 63
- Citations by year
- 2026: 4 · 2024: 2 · 2023: 2 · 2022: 1
- FWCI
- 0.44
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Current Research in Toxicology (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Morgridge Institute for Research
- U.S. Environmental Protection Agency (83573701)
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- co-culture, mouse embryonic fibroblasts, primary human hepatocytes, hepatocyte differentiation, xenobiotic metabolism, acetaminophen metabolism, hepatocyte dedifferentiation, hepatocyte expansion, drug metabolism, toxicity testing
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