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First published online December 9, 2005
Stem Cells Vol. 24 No. 3 March 2006, pp. 575 -586
doi:10.1634/stemcells.2005-0256; www.StemCells.com
© 2006 AlphaMed Press

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EMBRYONIC STEM CELLS

In Vitro Modeling of Paraxial and Lateral Mesoderm Differentiation Reveals Early Reversibility

Hidetoshi Sakuraia,b, Takumi Eraa, Lars Martin Jakta,c, Mitsuhiro Okadaa,c, Shigeru Nakaib, Satomi Nishikawaa, Shin-Ichi Nishikawaa

a Laboratory for Stem Cell Biology, RIKEN Center for Development Biology, Kobe, Japan;
b Science of In-Home Medicine, Health and Community Medicine, Nagoya University Graduate School of Medicine, Nagoya, Japan;
c Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, Japan Science and Technology Corporation, Discovery Research Laboratory, Tanabe Seiyaku Co., Ltd., Osaka, Japan

Key Words. Embryonic Stem cell • Mesoderm • Reversibility

Correspondence: Takumi Era, M.D., Ph.D., Laboratory for Stem Cell Biology, RIKEN Center for Development Biology, 2-2-3 Minatojimaminamimachi, Chuo-ku, Kobe, 650-0047, Japan. Telephone: 81-78-306-1893; Fax: 81-78-306-1895; e-mail: tera{at}cdb.riken.jp

Received June 7, 2005; accepted for publication September 6, 2005.
Endothelial cells (ECs) are thought to be derived mainly from the vascular endothelial growth factor receptor 2 (VEGFR-2)+ lateral mesoderm during early embryogenesis. In this study, we specified several pathways for EC differentiation using a murine embryonic stem (ES) cell differentiation culture system that is a model for cellular processes during early embryogenesis. Based on the results of in vitro fate analysis, we show that, in the main pathway, committed ECs are differentiated through the VEGFR-2+ platelet-derived growth factor receptor {alpha} (PDGFR-{alpha})– single-positive (VSP) population that is derived from the VEGFR-2+PDGFR-{alpha}+ double-positive (DP) population. This major differentiation course was also confirmed using DNA microarray analysis. In addition to this main pathway, however, ECs also can be generated from the VEGFR-2–PDGFR-{alpha}+ single-positive (PSP) population, which represents the paraxial mesodermal lineage and is also derived from the DP population. Our results strongly suggest that, even after differentiation from the common progenitor DP population into the VSP and PSP populations, these two populations continue spontaneous switching of their surface phenotype, which results in switching of their eventual fates. The rate of this interlineage conversion between VSP and PSP is unexpectedly high. Because of this potential to undergo fate switch, we conclude that ECs can be generated via multiple pathways in in vitro ES cell differentiation.




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