Efficient differentiation of human embryonic stem cells to definitive endoderm
Abstract
The potential of human embryonic stem (hES) cells to differentiate into cell types of a variety of organs has generated much excitement over the possible use of hES cells in therapeutic applications. Of great interest are organs derived from definitive endoderm, such as the pancreas. We have focused on directing hES cells to the definitive endoderm lineage as this step is a prerequisite for efficient differentiation to mature endoderm derivatives. Differentiation of hES cells in the presence of activin A and low serum produced cultures consisting of up to 80% definitive endoderm cells. This population was further enriched to near homogeneity using the cell-surface receptor CXCR4. The process of definitive endoderm formation in differentiating hES cell cultures includes an apparent epithelial-to-mesenchymal transition and a dynamic gene expression profile that are reminiscent of vertebrate gastrulation. These findings may facilitate the use of hES cells for therapeutic purposes and as in vitro models of development.
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CD184/CXCR4 was enriched in day-2 definitive endoderm in comparison to day-0 hPSCs and day-6 liver bud progenitors.
“Second, CD184/CXCR4 (a known definitive endoderm marker; D'Amour et al., 2005) was enriched in day-2 definitive endoderm in comparison to day-0 hPSCs and day-6 liver bud progenitors (Figure 3E; Table S1).”
From A Roadmap for Human Liver Differentiation from Pluripotent Stem Cells (2018) · cites this paper as D'Amour et al., 2005
Processes that govern early development of DE-derived tissues may be recapitulated during stem cell differentiation.
“Interest in the endoderm has intensified recently because processes that govern early development of DE-derived tissues may be recapitulated during stem cell differentiation [4,5]”
From A systematic screen for genes expressed in definitive endoderm by Serial Analysis of Gene Expression (SAGE) (2007) · cites this paper as [4]
Two groups achieved efficient differentiation of human and murine ES cells into DE by combining directive culture conditions and FACS sorting using CXCR4.
“More recently, two groups achieved efficient differentiation of human and murine ES cells into DE by combining directive culture conditions (serum concentration reduction and Activin supplements) and FACS sorting using the cell surface marker, CXCR4 [4,5,24].”
From A systematic screen for genes expressed in definitive endoderm by Serial Analysis of Gene Expression (SAGE) (2007) · cites this paper as [4]
A beta-like cell with behavior that fully recapitulates the physiology of a normal beta cell is yet to be realized.
“While a beta-like cell with behavior that fully recapitulates the physiology of a normal beta cell is yet to be realized”
From 100 years of insulin: celebrating the past, present and future of diabetes therapy (2021) · cites this paper as [135]
There has been a steady series of improvements to directed differentiation strategies for generating beta-like cells over the past 20 years.
“there has been a steady series of improvements to directed differentiation strategies over the past 20 years135–138.”
From 100 years of insulin: celebrating the past, present and future of diabetes therapy (2021) · cites this paper as [135]
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- Authors
- D'Amour, K. A., Agulnick, A. D., Eliazer, S., Kelly, O. G., Kroon, E., & Baetge, E. E.
- Corresponding author
- Baetge, Emmanuel E
- Venue
- Nature Biotechnology
- Published
- 2005-10-28
- Volume
- 23
- Issue
- 12
- Pages
- 1534-1541
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W2038404321
- Cited by
- 1,730
- References
- 3
- Citations by year
- 2026: 29 · 2025: 48 · 2024: 85 · 2023: 58 · 2022: 76 · 2021: 84 · 2020: 89 · 2019: 67 · 2018: 70 · 2017: 72 · 2016: 99 · 2015: 98 · 2014: 100 · 2013: 107 · 2012: 119
- FWCI
- 26.48
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- 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
- Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
- Keywords
- human embryonic stem cells, definitive endoderm differentiation, activin A, low serum culture, CXCR4, epithelial-mesenchymal transition, gastrulation, gene expression profiling, pancreatic development, in vitro organogenesis
- MeSH terms
- Animals, Cell Differentiation, Cells, Cultured, Endoderm, Humans, Stem Cells, Cell Culture Techniques, Tissue Engineering, Cell Proliferation, Mice
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