title: Differentiation of endothelial cells derived from mouse embryoid bodies: a possible in vitro vasculogenesis model.
abstract: Mouse embryonic stem cells (mES cells), which are pluripotent and self-renewal cells, are derived from the inner cell mass of mouse blastocysts. The objective of this study was to construct more efficient mES cell-derived embryoid bodies (EBs) for use as a vasculogenesis model and as an in vitro vascular toxicity testing model. EBs were formed for 3 days using hanging drop cultures and plated on gelatin-coated plates in endothelial growth medium-2 (EGM-2) to promote vascular development. The differentiation of mES cell-derived EBs was confirmed by reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry, and flow cytometry within 7 days after plating EBs. The mRNA and protein expressions of vascular endothelial growth factor receptors-2 (FLK-1), platelet endothelial cell adhesion molecule (PECAM), and vascular endothelial-cadherin (VE-cadherin) were observed in differentiated mES cells. When placed in matrigel, mES cell-derived endothelial like cells formed networks similar to vascular structures. mES cells were also exposed to 5-fluorouracil (5-FU), a strong inhibitor of vessel formation, and its cytotoxicity was determined using MTT assays. The inhibitory concentrations (IC50) of 5-FU for mES cells and C166 cells were 0.72 microM and 1.04 microM, respectively. These results demonstrate that mES cells can be used to study vasculogenesis and for cytotoxicity screening.
authors.affiliation: Department of Reproductive and Developmental Toxicology, National Institute of Toxicological Research, KFDA, Seoul 122-704, Republic of Korea.
authors.forename: Gi Dae
authors.initials: GD
authors.lastname: Kim
authors.forename: Gi Jin
authors.initials: GJ
authors.lastname: Kim
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Seok
authors.forename: Hyung-Min
authors.initials: HM
authors.lastname: Chung
authors.forename: Kew-Mahn
authors.initials: KM
authors.lastname: Chee
authors.forename: Gyu-Seek
authors.initials: GS
authors.lastname: Rhee
mesh_terms: D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D002196:Capillaries; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen; D003470:Culture Media; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000187:drug effects*; D042783:Endothelial Cells / Q000187:drug effects*; D005434:Flow Cytometry; D005472:Fluorouracil / Q000494:pharmacology; D007150:Immunohistochemistry; D007797:Laminin; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000235:genetics; D011509:Proteoglycans; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000964:Antimetabolites, Antineoplastic; D003470:Culture Media; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D012313:RNA; D003094:Collagen; D005472:Fluorouracil