Rapid generation of mature hepatocyte-like cells from human induced pluripotent stem cells by an efficient three-step protocol
Open accessAbstract
Liver transplantation is the only definitive treatment for end-stage cirrhosis and fulminant liver failure, but the lack of available donor livers is a major obstacle to liver transplantation. Recently, induced pluripotent stem cells (iPSCs) derived from the reprogramming of somatic fibroblasts, have been shown to resemble embryonic stem (ES) cells in that they have pluripotent properties and the potential to differentiate into all cell lineages in vitro , including hepatocytes. Thus, iPSCs could serve as a favorable cell source for a wide range of applications, including drug toxicity testing, cell transplantation, and patient-specific disease modeling. Here, we describe an efficient and rapid three-step protocol that is able to rapidly generate hepatocyte-like cells from human iPSCs. This occurs because the endodermal induction step allows for more efficient and definitive endoderm cell formation. We show that hepatocyte growth factor (HGF), which synergizes with activin A and Wnt3a, elevates the expression of the endodermal marker Foxa2 (forkhead box a2) by 39.3% compared to when HGF is absent (14.2%) during the endodermal induction step. In addition, iPSC-derived hepatocytes had a similar gene expression profile to mature hepatocytes. Importantly, the hepatocyte-like cells exhibited cytochrome P450 3A4 (CYP3A4) enzyme activity, secreted urea, uptake of low-density lipoprotein (LDL), and possessed the ability to store glycogen. Moreover, the hepatocyte-like cells rescued lethal fulminant hepatic failure in a nonobese diabetic severe combined immunodeficient mouse model. Conclusion : We have established a rapid and efficient differentiation protocol that is able to generate functional hepatocyte-like cells from human iPSCs. This may offer an alternative option for treatment of liver diseases. (Hepatology 2012)
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The PSC-to-liver bud differentiation system was significantly more rapid and yielded higher expression of liver bud markers compared to four extant liver differentiation methods.
“Finally, this PSC-to-liver bud differentiation system was significantly more rapid and yielded higher expression of liver bud markers, compared to four extant liver differentiation methods (Avior et al., 2015; Carpentier et al., 2016; Chen et al., 2012; Si-Tayeb et al., 2010; Zhao et al., 2013) (Figures 3A–3C and S3C).”
From A Roadmap for Human Liver Differentiation from Pluripotent Stem Cells (2018) · cites this paper as Chen et al., 2012
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“…generated by 4 methods: SR2 (the method described in the present work) and previously-reported methods (Carpentier et al., 2016; Avior et al., 2015; Chen et al., 2012) were stained for ALBUMIN (ALB) and Alpha1 anti-trypsin (AAT) with a DAPI nuclear counterstain and multiple fields were stitched…”
MethodSmall-Molecule-Driven Hepatocyte Differentiation of Human Pluripotent Stem Cells
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BackgroundMicroengineered Liver Tissues for Drug Testing
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- Authors
- Chen, Y.-F., Tseng, C.-Y., Wang, H.-W., Kuo, H.-C., Yang, V. W., & Lee, O. K.
- Affiliations
- Taipei Veterans General Hospital (TW)
- National Yang Ming University (TW)
- Genomics Research Center, Academia Sinica (TW)
- Stony Brook School (US)
- Corresponding author
- Lee, Oscar K.
- Venue
- Hepatology
- Published
- 2012-04
- Volume
- 55
- Issue
- 4
- Pages
- 1193-1203
- ISSN
- 0270-9139 (ISSN-L)
- PMC
- PMC3779307
- OpenAlex
- W2105782901
- Cited by
- 291
- References
- 1
- Citations by year
- 2026: 8 · 2025: 12 · 2024: 8 · 2023: 10 · 2022: 16 · 2021: 18 · 2020: 19 · 2019: 16 · 2018: 30 · 2017: 24 · 2016: 30 · 2015: 29 · 2014: 26 · 2013: 29 · 2012: 14
- FWCI
- 8.97
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Hepatology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Science Council (NSC100-2314-B-010-030-MY3, NSC 100, NSC99, NSC100-2120-M-010-001, 99-2911-I-009-101)
- University System of Taiwan
- Taipei Veterans General Hospital
- University of California, San Diego
- National Cancer Institute (R01CA084197)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK093680, R01DK052230)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Keywords
- hepatocyte-like cells, induced pluripotent stem cells, iPSC differentiation protocols, hepatocyte growth factor, FOXA2, activin A, Wnt3a, acute liver failure
- MeSH terms
- Animals, Carbon Tetrachloride, Cell Differentiation, Cells, Cultured, Disease Models, Animal, Glutamine, Humans, Mice, SCID, Treatment Outcome, Liver Failure, Hepatocyte Growth Factor, Cell Transplantation, Cell Lineage, Gene Expression Profiling, Hepatocytes, Pluripotent Stem Cells, Mice, Oncostatin M, In Vitro Techniques
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