Scalable production of tissue-like vascularized liver organoids from human PSCs
Open accessAbstract
The lack of physiological parity between 2D cell culture and in vivo culture has led to the development of more organotypic models, such as organoids. Organoid models have been developed for a number of tissues, including the liver. Current organoid protocols are characterized by a reliance on extracellular matrices (ECMs), patterning in 2D culture, costly growth factors and a lack of cellular diversity, structure, and organization. Current hepatic organoid models are generally simplistic and composed of hepatocytes or cholangiocytes, rendering them less physiologically relevant compared to native tissue. We have developed an approach that does not require 2D patterning, is ECM independent, and employs small molecules to mimic embryonic liver development that produces large quantities of liver-like organoids. Using single-cell RNA sequencing and immunofluorescence, we demonstrate a liver-like cellular repertoire, a higher order cellular complexity, presenting with vascular luminal structures, and a population of resident macrophages: Kupffer cells. The organoids exhibit key liver functions, including drug metabolism, serum protein production, urea synthesis and coagulation factor production, with preserved post-translational modifications such as N-glycosylation and functionality. The organoids can be transplanted and maintained long term in mice producing human albumin. The organoids exhibit a complex cellular repertoire reflective of the organ and have de novo vascularization and liver-like function. These characteristics are a prerequisite for many applications from cellular therapy, tissue engineering, drug toxicity assessment, and disease modeling to basic developmental biology.
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Liver organoids express key enzymes including members of the cytochrome P450 family and hepatic transporters, often at lower levels compared to native liver tissue.
“Studies have confirmed that liver organoids express key enzymes, including members of the cytochrome P450 (CYP450) family and hepatic transporters, though often at lower levels compared to native liver tissue14,15.”
From Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [15]
The Results sections on hepatic parenchymal cells and liver-like function report CYP2A6 expression and CYP1A2/CYP3A4 activity in the organoids. Our copy does not establish expression of hepatic transporters or show that these enzymes and transporters are generally expressed at lower levels than in native liver tissue; it reports some low-expression zonation markers, but that is not a native-liver comparison. No page numbers are available in our copy. (LLM, conf=0.85)
Experimental — accuracy not yet measuredProteomics can offer a temporal roadmap of cellular differentiation and functional development in organoids.
“By profiling protein expression dynamics, proteomics can offer a temporal roadmap of cellular differentiation and functional development in organoids14–17.”
From Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [15]
The Results section, “Proteomic analysis of iPSC-derived liver organoids” (page unavailable), uses protein profiles to identify liver-related functions and early developmental features. It compares organoids with primary liver, however, rather than profiling protein expression over time. The paper tracks differentiation across days using other measurements, so it does not establish the claimed temporal roadmap from proteomics. (LLM, conf=0.85)
Experimental — accuracy not yet measuredUnlike most prior studies, which typically analyze snapshots of organoid development at single timepoints, this study employs a longitudinal approach to track dynamic transitions of iHLO populations.
“Unlike most prior studies, which typically analyze snapshots of organoid development at single timepoints14,15, this study employs a longitudinal approach to track the dynamic transitions of iHLO populations.”
From Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [15]
The findings align with previous proteomic studies of liver organoids demonstrating their capacity to recapitulate liver-specific phenotypes.
“The findings from current study align with previous proteomic studies of liver organoids, which have demonstrated their capacity to recapitulate liver-specific phenotypes.”
From Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [15]
Harrison et al. compared iHLO protein profiles with native liver tissue and highlighted enrichment of detoxification pathways in both systems.
“Similarly, Harrison et al. compare iHLO protein profiles with native liver tissue and highlighted enrichment of detoxification pathways in both systems, emphasizing the relevance of organoids for metabolically intensive applications15.”
From Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [15]
Day 21 or around this timepoint has been identified as an important stage for liver organoid functional development in several previous studies.
“It is also noteworthy that Day 21, or around this timepoint, has been identified as an important stage for liver organoid functional development in several previous studies13,15,28,29.”
From Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [15]
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“It is also noteworthy that Day 21, or around this timepoint, has been identified as an important stage for liver organoid functional development in several previous studies 13,15,28,29 .”
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- Authors
- Harrison, S. P., Siller, R., Tanaka, Y., Chollet, M. E., de la Morena-Barrio, M. E., Xiang, Y., Patterson, B., Andersen, E., Bravo-Pérez, C., Kempf, H., Åsrud, K. S., Lunov, O., Dejneka, A., Mowinckel, M.-C., Stavik, B., Sandset, P. M., Melum, E., Baumgarten, S., Bonanini, F., Kurek, D., Mathapati, S., Almaas, R., Sharma, K., Wilson, S. R., Skottvoll, F. S., Boger, I. C., Bogen, I. L., Nyman, T. A., Wu, J. J., Bezrouk, A., Cizkova, D., Corral, J., Mokry, J., Zweigerdt, R., Park, I.-H., & Sullivan, G. J.
- Affiliations
- Oslo University Hospital (NO)
- University of Oslo (NO)
- Hôpital Maisonneuve-Rosemont (CA)
- Yale University (US)
- Medizinische Hochschule Hannover (DE)
- Czech Academy of Sciences (CZ)
- Mimetas (Netherlands) (NL)
- Durham University (GB)
- Centro Regional de Hemodonación (ES)
- Corresponding author
- Sullivan, Gareth J.
- ORCID
- Harrison, Sean P.
- Siller, Richard
- Tanaka, Yoshiaki
- Xiang, Yangfei
- Andersen, Elisabeth
- Kempf, Henning
- Lunov, Oleg
- Dejneka, Alexandr
- Stavik, Benedicte
- Sandset, Per Morten
- Melum, Espen
- Bonanini, Flavio
- Kurek, Dorota
- Mathapati, Santosh
- Almaas, Runar
- Sharma, Kulbhushan
- Wilson, Steven R.
- Skottvoll, Frøydis S.
- Bogen, Inger Lise
- Nyman, Tuula A.
- Wu, Jun Jie
- Corral, Javier
- Mokry, Jaroslav
- Zweigerdt, Robert
- Park, In-Hyun
- Sullivan, Gareth J.
- Venue
- Experimental & Molecular Medicine
- Published
- 2023-09-01
- Volume
- 55
- Issue
- 9
- Pages
- 2005-2024
- ISSN
- 1226-3613 (ISSN-L)
- 2092-6413
- PMC
- PMC10545717
- OpenAlex
- W4386318431
- Cited by
- 119
- References
- 81
- Citations by year
- 2026: 41 · 2025: 45 · 2024: 26 · 2023: 3 · 2022: 1 · 2021: 3
- FWCI
- 10.25
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Experimental & Molecular Medicine (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- St Andrews Research Repository (St Andrews Research Repository) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- NORA - Norwegian Open Research Archives (submittedVersion, cc-by)
- Funders
- Norges Forskningsråd (262613, 247624)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- liver organoids, vascularized organoids, human pluripotent stem cells, organotypic culture, embryonic liver development, Kupffer cells, drug metabolism, albumin synthesis, transplantation, N-glycosylation, cellular diversity, disease modeling
- MeSH terms
- Animals, Cells, Cultured, Humans, Liver, Organoids, Hepatocytes, Tissue Engineering, Mice
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