In Vitro Human Liver Model of Nonalcoholic Steatohepatitis by Coculturing Hepatocytes, Endothelial Cells, and Kupffer Cells
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The liver has a complex and unique microenvironment with multiple cell–cell interactions and internal vascular networks. Although nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease with multiple phases, no proper model could fully recapitulate the in vivo microenvironment to understand NAFLD progression. Here, an in vitro human liver model of NAFLD by coculturing human hepatocytes, umbilical vein endothelial cells (HUVECs), and Kupffer cells (KCs) into spheroids is presented. Analysis of indirect cross‐talk using conditioned media between steatotic spheroids—composed of hepatocellular carcinoma‐derived cells (HepG2) and HUVECs—and mouse KCs reveals that the latter can be activated showing increased cell area, elevated production of reactive oxygen species (ROS), and proinflammatory cytokines. Spheroids incorporating human KCs (HKCs) can also be induced into steatotic stage by supplementing fat. Steatotic spheroids with/without HKCs show different levels of steatotic stages through lipid accumulation and ROS production. Steatotic spheroids made from an immortalized hepatic progenitor cell line (HepaRG) compared to those made from HepG2 cells display similar trends of functionality, but elevated levels of proinflammatory cytokines, and improved reversibility of steatosis. The in vitro human liver system proposed makes strides in developing a model to mimic and monitor the progression of NAFLD.
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Previous liver models face the absence of lineage stage-specific paracrine signals from epithelial-mesenchymal cell-cell interactions.
“the absence of lineage stage-specific paracrine signals from epithelial-mesenchymal cell-cell interactions”
From Decellularized liver scaffolds for constructing drug-metabolically functional ex vivo human liver models (2025) · cites this paper as [5]
Previous liver models require a 3D environment to maintain cellular polarity and plasticity.
“the need for a 3D environment to maintain cellular maintaining polarity and plasticity”
From Decellularized liver scaffolds for constructing drug-metabolically functional ex vivo human liver models (2025) · cites this paper as [5]
Previous liver models can exhibit reduced activity levels relative to the liver's in vivo performance.
“It is observed that these models can exhibit reduced activity levels relative to the liver's in vivo performance, highlighting the need for further innovation in liver model development.”
From Decellularized liver scaffolds for constructing drug-metabolically functional ex vivo human liver models (2025) · cites this paper as [5]
Previous liver models may not fully capture the complexity of tissue-specific gene expression and liver functionality.
“Specifically, the models may not fully capture the complexity of tissue-specific gene expression and liver functionality [5,10].”
From Decellularized liver scaffolds for constructing drug-metabolically functional ex vivo human liver models (2025) · cites this paper as [5]
Previous liver models require a complex extracellular matrix that supports molecular, tissue, and cellular characteristics, metabolic regulation, and signal transduction.
“the requirement for a complex extracellular matrix (ECM) that support molecular, tissue, and cellular characteristics, metabolic regulation, and signal transduction [5,6]”
From Decellularized liver scaffolds for constructing drug-metabolically functional ex vivo human liver models (2025) · cites this paper as [5]
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Alikhani, H. K., Majidi, F., Ghasemzad, M., Shpichka, A. I., Nestorova, A., Hendi, Z., Rismani, E., Kuznetsova, D. S., Timashev, P., Vosough, M., et al. (2026) · Pharmacology Research & Perspectives · doi:10.1002/prp2.70306
Buffering cellular stress during mRNA delivery enables immunoregulatory interleukin-2 therapy
Chen, P., Hu, G., Nakashima, Y., Xu, Z., Hong, T., Chen, X., Kondo, S., Wolters, R., Yaginuma, H., Takasugi, S., et al. (2026) · Journal of Controlled Release · doi:10.1016/j.jconrel.2026.115393
- Authors
- Suurmond, C. E., Lasli, S., van den Dolder, F. W., Ung, A., Kim, H., Bandaru, P., Lee, K., Cho, H., Ahadian, S., Ashammakhi, N., Dokmeci, M. R., Lee, J., & Khademhosseini, A.
- Affiliations
- University of California, Los Angeles (US)
- Department of Bioengineering Henry Samueli School of Engineering and Applied Sciences University of California‐Los Angeles Los Angeles CA 90095 USA
- Corresponding author
- Lee, Junmin
- Khademhosseini, Ali
- ORCID
- Venue
- Advanced Healthcare Materials
- Publisher
- Wiley
- Published
- 2019-11-20
- Volume
- 8
- Issue
- 24
- Pages
- e1901379-e1901379
- ISSN
- 2192-2640 (ISSN-L)
- 2192-2659
- OpenAlex
- W2989983918
- Cited by
- 61
- References
- 31
- Citations by year
- 2026: 6 · 2025: 7 · 2024: 9 · 2023: 9 · 2022: 8 · 2021: 14 · 2020: 8
- FWCI
- 5.71
- Type
- journal article
- Language
- en
- Open access
- green
- dspace.library.uu.nl
- Licence
- Locations
- Advanced Healthcare Materials (publishedVersion)
- PubMed (publishedVersion)
- Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
- Infoscience (Ecole Polytechnique Fédérale de Lausanne) (submittedVersion, cc-by-nc-nd)
- University of Twente Research Information (submittedVersion)
- Funders
- National Institutes of Health (U01CA214411, R01GM126571)
- Topics
- Health Sciences › Medicine › Epidemiology › Liver Disease Diagnosis and Treatment
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- nonalcoholic steatohepatitis, non-alcoholic fatty liver disease, in vitro liver models, spheroids, Kupffer cells, hepatocytes, umbilical vein endothelial cells, steatosis, lipid accumulation, reactive oxygen species, inflammatory cytokines, HepaRG cells, HepG2 cells, hepatic progenitor cells, disease progression
- MeSH terms
- Humans, Kupffer Cells, Culture Media, Conditioned, Reactive Oxygen Species, Hepatocytes, Endothelial Cells, Hep G2 Cells, Human Umbilical Vein Endothelial Cells, Non-alcoholic Fatty Liver Disease
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