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A system to enrich for primitive streak-derivatives, definitive endoderm and mesoderm, from pluripotent cells in culture


Vassilieva, S and Goh, HN and Lau, KX and Hughes, JN and Familari, M and Rathjen, PD and Rathjen, J, A system to enrich for primitive streak-derivatives, definitive endoderm and mesoderm, from pluripotent cells in culture, PLoS One, 7, (6) Article e38645. ISSN 1932-6203 (2012) [Refereed Article]


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Licensed under Creative Commons Attribution 2.5 Generic (CC BY 2.5)

DOI: doi:10.1371/journal.pone.0038645


Two lineages of endoderm develop during mammalian embryogenesis, the primitive endoderm in the pre-implantation blastocyst and the definitive endoderm at gastrulation. This complexity of endoderm cell populations is mirrored during pluripotent cell differentiation in vitro and has hindered the identification and purification of the definitive endoderm for use as a substrate for further differentiation. The aggregation and differentiation of early primitive ectoderm-like (EPL) cells, resulting in the formation of EPL-cell derived embryoid bodies (EPLEBs), is a model of gastrulation that progresses through the sequential formation of primitive streak-like intermediates to nascent mesoderm and more differentiated mesoderm populations. EPL cell-derived EBs have been further analysed for the formation of definitive endoderm by detailed morphological studies, gene expression and a protein uptake assay. In comparison to embryoid bodies derived from ES cells, which form primitive and definitive endoderm, the endoderm compartment of embryoid bodies formed from EPL cells was comprised almost exclusively of definitive endoderm. Definitive endoderm was defined as a population of squamous cells that expressed Sox17, CXCR4 and Trh, which formed without the prior formation of primitive endoderm and was unable to endocytose horseradish peroxidase from the medium. Definitive endoderm formed in EPLEBs provides a substrate for further differentiation into specific endoderm lineages; these lineages can be used as research tools for understanding the mechanisms controlling lineage establishment and the nature of the transient intermediates formed. The similarity between mouse EPL cells and human ES cells suggests EPLEBs can be used as a model system for the development of technologies to enrich for the formation of human ES cell-derived definitive endoderm in the future.

Item Details

Item Type:Refereed Article
Keywords:embryonic stem cell, differentiation, primitive streak, visceral endoderm, primitive endoderm, definitive endoderm
Research Division:Biological Sciences
Research Group:Biochemistry and cell biology
Research Field:Cell development, proliferation and death
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the biological sciences
UTAS Author:Rathjen, PD (Professor Peter Rathjen)
UTAS Author:Rathjen, J (Dr Joy Rathjen)
ID Code:78691
Year Published:2012
Web of Science® Times Cited:5
Deposited By:Menzies Institute for Medical Research
Deposited On:2012-07-19
Last Modified:2019-04-15
Downloads:450 View Download Statistics

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