Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen
Open accessAbstract
The establishment of protocols to differentiate human pluripotent stem cells (hPSCs) including embryonic (ESC) and induced pluripotent (iPSC) stem cells into functional hepatocyte-like cells (HLCs) creates new opportunities to study liver metabolism, genetic diseases and infection of hepatotropic viruses (hepatitis B and C viruses) in the context of specific genetic background. While supporting efficient differentiation to HLCs, the published protocols are limited in terms of differentiation into fully mature hepatocytes and in a smaller-well format. This limitation handicaps the application of these cells to high-throughput assays. Here we describe a protocol allowing efficient and consistent hepatic differentiation of hPSCs in 384-well plates into functional hepatocyte-like cells, which remain differentiated for more than 3weeks. This protocol affords the unique opportunity to miniaturize the hPSC-based differentiation technology and facilitates screening for molecules in modulating liver differentiation, metabolism, genetic network, and response to infection or other external stimuli.
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HNF4A+ liver progenitors were typically formed on later days (by day 10-14) in other differentiation protocols.
“whereas HNF4A+ liver progenitors were typically formed on later days (by day 10–14) in other differentiation protocols (Carpentier et al., 2016; Hay et al., 2008; Si-Tayeb et al., 2010; Zhao et al., 2013).”
From A Roadmap for Human Liver Differentiation from Pluripotent Stem Cells (2018) · cites this paper as Carpentier et al., 2016
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 Carpentier et al., 2016
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A Roadmap for Human Liver Differentiation from Pluripotent Stem Cells
Ang, L. T., Tan, A. K. Y., Autio, M. I., Goh, S. H., Choo, S. H., Lee, K. L., Tan, J., Pan, B., Lee, J. J. H., Lum, J. J., et al. (2018) · Cell Reports · doi:10.1016/j.celrep.2018.01.087 · Cited by 220 · see citing works
“Finally, the cells were washed once with 100 ng/mL DAPI/PBS once and twice with 1x PBS before visualization using Zeiss Axio Vision.”
BackgroundMethodResultInfluential
- Authors
- Carpentier, A., Nimgaonkar, I., Chu, V., Xia, Y., Hu, Z., & Liang, T. J.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Carpentier, Arnaud
- Liang, T. Jake
- Venue
- Stem Cell Research
- Publisher
- Elsevier BV
- Published
- 2016-05
- Volume
- 16
- Issue
- 3
- Pages
- 640-650
- ISSN
- 1873-5061 (ISSN-L)
- PMC
- PMC4903911
- OpenAlex
- W2316384098
- Cited by
- 90
- References
- 1
- Citations by year
- 2026: 7 · 2025: 8 · 2024: 5 · 2023: 5 · 2022: 13 · 2021: 11 · 2020: 14 · 2019: 9 · 2018: 9 · 2017: 6 · 2016: 3
- FWCI
- 4.68
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Stem Cell Research (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- HZI (Helmholtz Centre for Infection Research) (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- NIDDK (Z01 DK54504, DK54505)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Keywords
- human pluripotent stem cells, hepatocyte-like cells, hepatic differentiation, 384-well plate, high-throughput screening, induced pluripotent stem cells, embryonic stem cells, hepatic metabolism, hepatotropic viruses, hepatitis B virus, hepatitis C virus
- MeSH terms
- alpha-Fetoproteins, Cell Differentiation, Humans, Microscopy, Fluorescence, Miniaturization, Hepatocytes, Pluripotent Stem Cells, Octamer Transcription Factor-3, Cytochrome P-450 CYP3A, Hepatocyte Nuclear Factor 4
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