Characterization of mesodermal cells in in vitro ES cell differentiation

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 16606005
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 幹細胞生物学
• Research Institution: RIKEN
• Principal Investigator: ERA Takumi RIKEN, Stem cell biology Laboratory, Researcher, 幹細胞研究グループ, 研究員 (00273706)
• Project Period (FY): 2004 – 2005
• Keywords: ES cell / In vitro differentiation / mesoderm / PDGFR / VEGFR2

Research Abstract

Mesoderm cells segregate from primitive streak and diversified into three major population including paraxial, lateral and intermediate type. We attempted to visualize and isolate these types of mesodermal cells in ES cell culture using two types of surface markers, PDGFRα and VEGFR2. Based on the expression pattern of the markers, ES cell-derived mesodermal cells are divided into three population composed of PDGFRα^+, VEGFR2^+(DP), PDGFRα^+, VEGFR2^-(PSP) and PDGFRα^-, VEGFR2^+(VSP). Fate and gene expression analyses have revealed that PSP and VSP show the characteristics of paraxial and lateral mesoderm, respectively. DP could give rise to both PSP and VSP, suggesting that DP contained a common precursor for PSP and VSP. Early gastrula organizer can contribute to another type of mesoderm, axial mesoderm. Goosecoid (Gsc) is a transcriptional factor carrying homeobox and is a ideal candidate of organizer's markers as it is specifically expressed in organizer region during embryogenesis. To visualize the ES cell-derived Gsc-expressing cells, we have generated goosecoid knock-in ES cell using eGFP reporter gene. We tried to establish a suitable condition in the absence of serum and finally found a good culture condition in which almost all of differentiated ES cells were expressing Gsc. Along with the differentiation, Gsc-GFP positive cells were clearly separated into two major populations by the pattern of E-cadherin(ECD) expression. ECD^+ population in Gsc^+ shows endodermal phenotype by cell fate, gene expression and cell morphology. Another population that does not express ECD corresponds to mesoderm lineage and gives rise to the mesodermal descendants such as bone and endothelial cells. A single cell analysis of Gsc^+ cells has identified the mesendodermal progenitors in ES cell culture, which can give rise to endoderm and mesoderm. The current study also has the potential to be useful for understanding molecular event involved in mesodermal differentiation.

Research Products

• [Journal Article] In vitro modeling of paraxial and lateral mesoderm differentation reveals early reversibility (2006)
  • Author(s): Sakurai, H. et al.
  • Journal Title: Stem Cells 24 Pages: 575-586
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Microarray analysis of PDGFRα? populations in ES cell differentiation culture identifies genes involved in differentiation of mesoderm and mesenchyme in cluding ARID 3d that is essential for development of embryonic mesenchymal cells. (2006)
  • Author(s): Takebe, A. et al.
  • Journal Title: Devwlopmental Biology 293 Pages: 25-37
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Microarray analysis of PDGFRα^+ populations in ES cell differentiation culture identifies genes involved in differentiation of mesoderm and mesenchyme including ARID3b that is essential for development of embryonic mesenchymal cells. (2006)
  • Author(s): Takebe, A., Era, T., Okada, M., Jakt, LM., Kuroda, Y., Nishikawa, S-I.
  • Journal Title: Dev.Biol. 293 Pages: 25-37
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] In vitro modeling of paraxial and lateral mesoderm differentiation reveals early reversibility. (2006)
  • Author(s): Sakurai, H., Era, T., Jakt, LM., Okada, M., Nakai, S., Nishikawa, S., Nishikawa, S-I.
  • Journal Title: Stem Cells 24 Pages: 575-586
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells. (2005)
  • Author(s): Yasunaga, M. et al.
  • Journal Title: Nature Biotech 23 Pages: 1542-1550
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of mesendoderm, a diverging point of the definitive endoderm and mesoderm in embryonic stem cell differetiation culture. (2005)
  • Author(s): Tada, S. et al.
  • Journal Title: Development 132 Pages: 4363-4374
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Notch signals inhibit the development of erythroid/megakaryocytic cells by suppressing GATA-1 activity through the induction of HES1. (2005)
  • Author(s): Ishiko, E. et al.
  • Journal Title: J. Biol. Chem. 280 Pages: 4929-4939
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells. (2005)
  • Author(s): Yasunaga, M., Tada, S., Torikai-Nishikawa, S., Nakano, Y., Okada, M., Yakt, L.M., Nishikawa, S., Chiba, T., Era, T., Nishikawa, S-I.
  • Journal Title: Nature Biotech 23 Pages: 1542-1550
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of mesendoderm, a diverging point of the definitive endoderm and mesoderm in embryonic stem cell differentiation culture. (2005)
  • Author(s): Tada, S., Era, T., Furusawa, C., Sakurai, H., Nishikawa, S., Kinoshita, M., Nakao, K., Chiba, T., Nishikawa, S-I.
  • Journal Title: Development 132 Pages: 4363-4374
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Notch signals inhibit the development of erythroid/megakaryocytic cells by suppressing GATA-1 activity through the induction of HES1. (2005)
  • Author(s): Ishiko, E., Matsumura, I., Ezoe, S., Gale, K., Ishiko, J., Satoh, Y., Tanaka, H., Shibayama, H., Mizuki M., Era, T., Enver, T., Kanakura, Y.
  • Journal Title: J.Biol.Chem. 280 Pages: 4929-4939
  • Description: 「研究成果報告書概要(欧文)」より