title: Characterization of mesendoderm: a diverging point of the definitive endoderm and mesoderm in embryonic stem cell differentiation culture.
abstract: 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)+ PDGFRalpha(alphaR)+ population and is selectively induced from ES cells under defined conditions containing either activin or nodal. Subsequently, it diverges to Gsc+ ECD+ alphaR- and Gsc+ ECD- alphaR+ intermediates that eventually differentiate into definitive endoderm and mesodermal lineages, respectively. The presence of mesendodermal cells in nascent Gsc+ ECD+ alphaR+ 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.
authors.affiliation: Laboratory for Stem Cell Biology, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, 650-0047, Japan.
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Tada
authors.forename: Takumi
authors.initials: T
authors.lastname: Era
authors.forename: Chikara
authors.initials: C
authors.lastname: Furusawa
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Sakurai
authors.forename: Satomi
authors.initials: S
authors.lastname: Nishikawa
authors.forename: Masaki
authors.initials: M
authors.lastname: Kinoshita
authors.forename: Kazuki
authors.initials: K
authors.lastname: Nakao
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Chiba
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Nishikawa
mesh_terms: D028341:Activins / Q000502:physiology; D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage; D016895:Culture Media, Serum-Free; D004707:Endoderm / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D051956:Goosecoid Protein / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology*; D055457:Nodal Protein; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000502:physiology
chemical_list: D015820:Cadherins; D016895:Culture Media, Serum-Free; D051956:Goosecoid Protein; C497993:Gsc protein, mouse; D055457:Nodal Protein; C526008:Nodal protein, mouse; D011993:Recombinant Fusion Proteins; D016212:Transforming Growth Factor beta; D028341:Activins; D049452:Green Fluorescent Proteins; D020796:Receptor, Platelet-Derived Growth Factor alpha