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First published online 1 September 2005
doi: 10.1242/dev.02005
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1 Laboratory for Stem Cell Biology, RIKEN Center for Developmental Biology,
2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, 650-0047, Japan
2 Department of Gastroenterology and Hepatology, Graduate school of Medicine,
Kyoto University, 54 Shogoinkawara-cho, Sakyo-ku, Kyoto, 606-8507, Japan
3 Department of Bioinformatic Engineering, Graduate School of Information
Science and Technology, Osaka University, 2-1 Yamada-oka, Suita City, Osaka,
565-0871, Japan
4 Laboratory for Animal Resources and Genetic Engineering, RIKEN Center for
Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, 650-0047,
Japan
* Author for correspondence (e-mail: tera{at}cdb.riken.jp)
Accepted 19 July 2005
Bipotent mesendoderm that can give rise to both endoderm and mesoderm is an
established entity from C. elegans to zebrafish. Although previous
studies in mouse embryo indicated the presence of bi-potent mesendoderm cells
in the organizer region, characterization of mesendoderm and its
differentiation processes are still unclear. As bi-potent mesendoderm is
implicated as the major precursor of definitive endoderm, its identification
is also essential for exploring the differentiation of definitive endoderm. In
this study, we have established embryonic stem (ES) cell lines that carry
GFP gene in the goosecoid (Gsc) gene locus and have
investigated the differentiation course of mesendodermal cells using Gsc
expression as a marker. Our results show that mesendoderm is represented as a
Gsc-GFP+E-cadherin(ECD)+PDGFR
(R)+
population and is selectively induced from ES cells under defined conditions
containing either activin or nodal. Subsequently, it diverges to
Gsc+ECD+R- and
Gsc+ECD-R+ intermediates that
eventually differentiate into definitive endoderm and mesodermal lineages,
respectively. The presence of mesendodermal cells in nascent
Gsc+ECD+R+ population was also
confirmed by single cell analysis. Finally, we show that the defined culture
condition and surface markers developed in this study are applicable for
obtaining pure mesendodermal cells and their immediate progenies from
genetically unmanipulated ES cells.
Key words: ES cell, Mesendoderm, Goosecoid, Endoderm, Mouse
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