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Inflammatory Cytokine TNFα Promotes the Long-Term Expansion of Primary Hepatocytes in 3D Culture
Open accessAbstract
In the healthy adult liver, most hepatocytes proliferate minimally. However, upon physical or chemical injury to the liver, hepatocytes proliferate extensively in vivo under the direction of multiple extracellular cues, including Wnt and pro-inflammatory signals. Currently, liver organoids can be generated readily in vitro from bile-duct epithelial cells, but not hepatocytes. Here, we show that TNFα, an injury-induced inflammatory cytokine, promotes the expansion of hepatocytes in 3D culture and enables serial passaging and long-term culture for more than 6 months. Single-cell RNA sequencing reveals broad expression of hepatocyte markers. Strikingly, in vitro-expanded hepatocytes engrafted, and significantly repopulated, the injured livers of Fah-/- mice. We anticipate that tissue repair signals can be harnessed to promote the expansion of otherwise hard-to-culture cell-types, with broad implications.
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“Organoids of murine intestines31, livers35,125,126 and pancreas69,146 were successfully transplanted into mice, and the transplanted organoids could restore organ dysfunction.”
From Organoids (2022) · cites this paper as [125]
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“In the case of the liver, both cholangiocyte (ductal cell of the bile duct)35,36 and hepatocyte organoids125,126 have been established, both from mouse and human tissue samples.”
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“In the example of hepatocyte organoids125, hepatocyte functions are preserved, yet the liver tissue architecture does not match the native tissue where hepatocytes are arranged in cords.”
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“Several of the most dramatic expansion results reported to date—including near-unlimited theoretical fold-expansion and multi-month clonal proliferation—have been demonstrated almost exclusively in mouse hepatocytes [18,31], whereas human systems have generally achieved more modest expansion with measurable trade-offs in redifferentiation and repopulation capacity at higher expansion levels [4,17]”
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“Compact 3D liver models enhanced with tumor necrosis factor-α (TNFα) have demonstrated the capacity to sustain long-term hepatocyte culture, encompassing serial passaging and expansion over several months, and thus illustrate the importance of precisely regulated inflammatory signaling in facilitating hepatocyte plasticity and viability [31].”
From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [31]
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Metadata
- Authors
- Peng, W. C., Logan, C. Y., Fish, M., Anbarchian, T., Aguisanda, F., Álvarez-Varela, A., Wu, P., Jin, Y., Zhu, J., Li, B., Grompe, M., Wang, B., & Nusse, R.
- Affiliations
- Howard Hughes Medical Institute (US)
- Stanford University (US)
- Oregon Health & Science University (US)
- University of California, San Francisco (US)
- Corresponding author
- Peng, Weng Chuan
- ORCID
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2018-11
- Volume
- 175
- Issue
- 6
- Pages
- 1607-1619.e15
- ISSN
- 0092-8674 (ISSN-L)
- PMC
- PMC6497386
- OpenAlex
- W2902626180
- Cited by
- 331
- References
- 78
- Citations by year
- 2026: 33 · 2025: 43 · 2024: 48 · 2023: 40 · 2022: 49 · 2021: 59 · 2020: 39 · 2019: 19 · 2018: 1
- FWCI
- 18.63
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- California Institute for Regenerative Medicine
- NIH (S10OD020141)
- Stanford Child Health Research Institute
- Burroughs Wellcome Fund
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- TNF-α, hepatocyte expansion, primary hepatocytes, 3D cell culture, liver regeneration, hepatocyte engraftment, liver injury, hepatocyte markers, long-term culture
- MeSH terms
- Animals, Antigens, Differentiation, Cell Line, Transformed, Humans, Liver, Time Factors, Tumor Necrosis Factor-alpha, Mice, Knockout, Spheroids, Cellular, Cell Culture Techniques, Hepatocytes, Cell Proliferation, Hep G2 Cells, Human Umbilical Vein Endothelial Cells
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