Generation of multi-cellular human liver organoids from pluripotent stem cells
Open accessAbstract
A growing number of in vitro hepatic models exist to study human genetics, liver biology, disease modeling and drug development and range from 2D hepatocytes to 3D multi-cellular tissues that are derived from human stem cells. However, stem cell-based models generally suffer from batch-, clone- and donor-dependent variability, hindering broader usage in diverse biomedical applications. To circumvent this challenge, we herein describe a reproducible protocol to generate human liver organoids in 20-25 days derived from pluripotent stem cells (PSCs). These organoids are intra-luminally polarized to form canalicular structures and are comprised of mainly hepatic epithelial cells, co-differentiated with stellate-like and hepatic macrophage-like cells that enables hepatic inflammatory disease modeling in vitro. These multi-lineage liver organoids express hepatocyte genes, secrete albumin and have vital metabolic functions. This protocol utilizes PSC derived 3D human liver organoids as a renewable, reproducible and personalized cell source, thus facilitating disease modeling and mechanistic studies with a future goal of developing novel therapeutics against currently intractable diseases.
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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 [29]
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Morozumi, R., Kinoshita, M., Takahashi, R., Mishima, M., Izawa, K., Sugiyama, K.-I., & Tsuda, M. (2026) · Biology Methods and Protocols · doi:10.1093/biomethods/bpag036
Simone, U. D., Locatelli, C., Caloni, F., Lonati, D., Russo, L. A., & Coccini, T. (2026) · Neurotoxicity Research · doi:10.1007/s12640-026-00808-z
- Authors
- Thompson, W. L., & Takebe, T.
- Affiliations
- Cincinnati Children's Hospital Medical Center (US)
- Tokyo Medical and Dental University (JP)
- Corresponding author
- Takanori Takebe
- Venue
- Methods in Cell Biology
- Publisher
- Elsevier
- Published
- 2020
- Volume
- 159
- Pages
- 47-68
- ISSN
- 0091-679X (ISSN-L)
- PMC
- PMC8011941
- OpenAlex
- W3027429375
- Cited by
- 55
- References
- 32
- Citations by year
- 2026: 9 · 2025: 11 · 2024: 10 · 2023: 13 · 2022: 9 · 2021: 3
- FWCI
- 8.46
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Methods in cell biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Japan Society for the Promotion of Science (18H02800, 19K22416)
- National Institute of Diabetes and Digestive and Kidney Diseases (DP2DK128799, UH3DK119982, UG3DK119982, P30DK078392)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- liver organoids, pluripotent stem cells, 3D organoid culture, hepatic stellate cells, hepatic macrophages, hepatocyte differentiation, drug development
- MeSH terms
- Cell Differentiation, Fatty Acids, Humans, Liver, Organoids, Cell Culture Techniques, Pluripotent Stem Cells
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