Efficient endoderm induction from human pluripotent stem cells by logically directing signals controlling lineage bifurcations
Open accessAbstract
Human pluripotent stem cell (hPSC) differentiation typically yields heterogeneous populations. Knowledge of signals controlling embryonic lineage bifurcations could efficiently yield desired cell types through exclusion of alternate fates. Therefore, we revisited signals driving induction and anterior-posterior patterning of definitive endoderm to generate a coherent roadmap for endoderm differentiation. With striking temporal dynamics, BMP and Wnt initially specified anterior primitive streak (progenitor to endoderm), yet, 24 hr later, suppressed endoderm and induced mesoderm. At lineage bifurcations, cross-repressive signals separated mutually exclusive fates; TGF-β and BMP/MAPK respectively induced pancreas versus liver from endoderm by suppressing the alternate lineage. We systematically blockaded alternate fates throughout multiple consecutive bifurcations, thereby efficiently differentiating multiple hPSC lines exclusively into endoderm and its derivatives. Comprehensive transcriptional and chromatin mapping of highly pure endodermal populations revealed that endodermal enhancers existed in a surprising diversity of "pre-enhancer" states before activation, reflecting the establishment of a permissive chromatin landscape as a prelude to differentiation.
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These lineage segregations could be efficiently negotiated by providing the relevant inductive signal(s) to drive differentiation toward the desired lineage while repressing the signal(s) that otherwise promoted the alternate fate.
“With this knowledge, these lineage segregations could be efficiently negotiated by providing the relevant inductive signal(s) to drive differentiation toward the desired lineage while repressing the signal(s) that otherwise promoted the alternate fate (Loh et al., 2014).”
From A Roadmap for Human Liver Differentiation from Pluripotent Stem Cells (2018) · cites this paper as Loh et al., 2014
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- Authors
- Loh, K. M., Ang, L. T., Zhang, J., Kumar, V., Ang, J., Auyeong, J. Q., Lee, K. L., Choo, S. H., Lim, C. Y. Y., Nichane, M., Tan, J., Noghabi, M. S., Azzola, L., Ng, E. S., Durruthy-Durruthy, J., Sebastiano, V., Poellinger, L., Elefanty, A. G., Stanley, E. G., Chen, Q., Prabhakar, S., Weissman, I. L., & Lim, B.
- Affiliations
- Agency for Science, Technology and Research (SG)
- National University of Singapore (SG)
- Monash University (AU)
- Royal Children's Hospital (AU)
- Institute for Stem Cell Biology and Regenerative Medicine (US)
- Corresponding author
- Loh, Kyle M.
- Ang, Lay Teng
- Lim, Bing
- ORCID
- Venue
- Cell Stem Cell
- Publisher
- Elsevier BV
- Published
- 2014-02
- Volume
- 14
- Issue
- 2
- Pages
- 237-252
- ISSN
- 1875-9777 (ISSN-L)
- 1934-5909
- PMC
- PMC4045507
- OpenAlex
- W2074884450
- Cited by
- 418
- References
- 7
- Citations by year
- 2026: 34 · 2025: 37 · 2024: 36 · 2023: 21 · 2022: 55 · 2021: 43 · 2020: 43 · 2019: 41 · 2018: 25 · 2017: 24 · 2016: 31 · 2015: 17 · 2014: 10
- FWCI
- 11.21
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell stem cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- National University of Singapore (submittedVersion)
- Funders
- National Heart, Lung, and Blood Institute (U01HL099995, U01HL099999)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Keywords
- human pluripotent stem cells, BMP signaling, Wnt signaling, TGF-beta signaling, MAPK signaling, liver differentiation, anterior-posterior patterning, primitive streak, cell fate specification, chromatin accessibility
- MeSH terms
- Animals, Base Sequence, Chromatin, Digestive System, Endoderm, Enhancer Elements, Genetic, Fibroblast Growth Factors, Humans, Liver, Molecular Sequence Data, Pancreas, Protein Binding, Transcription, Genetic, Signal Transduction, Transforming Growth Factor beta, Culture Media, Serum-Free, Cell Lineage, Bone Morphogenetic Proteins, Body Patterning, MAP Kinase Signaling System, Pluripotent Stem Cells, Epigenesis, Genetic, Wnt Proteins, Mice, Embryonic Stem Cells, Primitive Streak
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