Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells
Abstract
Preparation of specific lineages at high purities from embryonic stem (ES) cells requires both selective culture conditions and markers to guide and monitor the differentiation. In this study, we distinguished definitive and visceral endoderm by using a mouse ES cell line that bears the gfp and human IL2R alpha (also known as CD25) marker genes in the goosecoid (Gsc) and Sox17 loci, respectively. This cell line allowed us to monitor the generation of Gsc+ Sox17+ definitive endoderm and Gsc- Sox17+ visceral endoderm and to define culture conditions that differentially induce definitive and visceral endoderm. By comparing the gene expression profiles of definitive and visceral endoderm, we identified seven surface molecules that are expressed differentially in the two populations. One of the seven markers, Cxcr4, to which a monoclonal antibody is available allowed us to monitor and purify the Gsc+ population from genetically unmanipulated ES cells under the condition that selects definitive endoderm.
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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 [5]
CXCR4 is not an ideal marker for DE as it is widely expressed in the gastrulation stage mouse embryo.
“Although useful, CXCR4 is not an ideal marker for the DE as it is widely expressed in the gastulation stage mouse embryo (Table 1 and [5,25]).”
From A systematic screen for genes expressed in definitive endoderm by Serial Analysis of Gene Expression (SAGE) (2007) · cites this paper as [5]
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 [5]
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Generation of Functional Human Hepatic Endoderm From Human Induced Pluripotent Stem Cells
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Definitive endoderm of the mouse embryo: Formation, cell fates, and morphogenetic function
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Anterior definitive endoderm from ESCs reveals a role for FGF signaling
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“Although useful, CXCR4 is not an ideal marker for the DE as it is widely expressed in the gastulation stage mouse embryo (Table 1 and [5,25]).”
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- Authors
- Yasunaga, M., Tada, S., Torikai-Nishikawa, S., Nakano, Y., Okada, M., Jakt, L. M., Nishikawa, S., Chiba, T., Era, T., & Nishikawa, S.-I.
- Affiliations
- Kyoto University (JP)
- RIKEN (JP)
- Venue
- Nature Biotechnology
- Published
- 2005-11-27
- Volume
- 23
- Issue
- 12
- Pages
- 1542-1550
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W1965419330
- Cited by
- 471
- References
- 1
- Citations by year
- 2025: 5 · 2024: 4 · 2023: 7 · 2022: 9 · 2021: 7 · 2020: 9 · 2019: 4 · 2018: 11 · 2017: 12 · 2016: 22 · 2015: 28 · 2014: 31 · 2013: 35 · 2012: 35
- FWCI
- 15.20
- 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
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- definitive endoderm differentiation, mouse embryonic stem cells, goosecoid, SOX17, CD25, GFP reporter, CXCR4, lineage markers, gene expression profiling, cell purification, culture conditions
- MeSH terms
- Animals, Cell Differentiation, Cells, Cultured, Endoderm, Stem Cells, Cell Culture Techniques, Tissue Engineering, Cell Proliferation, Mice
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