title: Differentiation of lymphatic endothelial cells from embryonic stem cells on OP9 stromal cells.
abstract: OBJECTIVE
  The discovery of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 (VEGFR-3) has started to provide an understanding of the molecular mechanisms of lymphangiogenesis. The homeobox gene prox1 has been proven to specify lymphatic endothelial cells (ECs) from blood ECs. We investigated the process of lymphatic EC (LEC) differentiation using embryonic stem (ES) cells.
  
  
  RESULTS
  VEGFR-2+ cells derived from ES cells differentiated into LECs at day 3 on OP9 stromal cells defined by the expression of prox1, VEGFR-3, and another lymphatic marker podoplanin. VEGFR-2+ cells gave rise to LYVE-1+ embryonic ECs, which were negative for prox1 on day 1 but turned to prox1+ LECs by day 3. VEGFR-3-Fc or Tie2-Fc, sequestering VEGF-C or angiopoietin1 (Ang1), suppressed colony formation of LECs on OP9 cells. However, addition of VEGF-C and Ang1 in combination with VEGF to the culture of VEGFR-2+ cells on collagen-coated dishes failed to induce LECs. LEC-inducing activity of OP9 cells was fully reproduced on paraformaldehyde-fixed OP9 cells with the conditioned medium.
  
  
  CONCLUSIONS
  We succeeded in differentiating LECs from ES cells and revealed the requirements of VEGF-C, Ang1, and other unknown factors for LEC differentiation.
authors.affiliation: Molecular and Cancer Research Unit, HMRO, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Kono
authors.forename: Hajime
authors.initials: H
authors.lastname: Kubo
authors.forename: Chikashi
authors.initials: C
authors.lastname: Shimazu
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Ueda
authors.forename: Meiko
authors.initials: M
authors.lastname: Takahashi
authors.forename: Kentoku
authors.initials: K
authors.lastname: Yanagi
authors.forename: Naoya
authors.initials: N
authors.lastname: Fujita
authors.forename: Takashi
authors.initials: T
authors.lastname: Tsuruo
authors.forename: Hiromi
authors.initials: H
authors.lastname: Wada
authors.forename: Jun K
authors.initials: JK
authors.lastname: Yamashita
mesh_terms: D042683:Angiopoietin-1 / Q000494:pharmacology; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004338:Drug Combinations; D004622:Embryo, Mammalian / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D051379:Mice; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology*; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D042582:Vascular Endothelial Growth Factor C / Q000494:pharmacology / Q000502:physiology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D042683:Angiopoietin-1; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D042461:Vascular Endothelial Growth Factor A; D042582:Vascular Endothelial Growth Factor C; D040301:Vascular Endothelial Growth Factor Receptor-2