title: Osteogenic differentiation of murine embryonic stem cells is mediated by fibroblast growth factor receptors.
abstract: The mechanisms involved in the control of embryonic stem (ES) cell differentiation are yet to be fully elucidated. However, it has become clear that the family of fibroblast growth factors (FGFs) are centrally involved. In this study we examined the role of the FGF receptors (FGFRs 1-4) during osteogenesis in murine ES cells. Single cells were obtained after the formation of embryoid bodies, cultured on gelatin-coated plates, and coaxed to differentiate along the osteogenic lineage. Upregulation of genes was analyzed at both the transcript and protein levels using gene array, relative-quantitative PCR (RQ-PCR), and Western blotting. Deposition of a mineralized matrix was evaluated with Alizarin Red staining. An FGFR1-specific antibody was generated and used to block FGFR1 activity in mES cells during osteogenic differentiation. Upon induction of osteogenic differentiation in mES cells, all four FGFRs were clearly upregulated at both the transcript and protein levels with a number of genes known to be involved in osteogenic differentiation including bone morphogenetic proteins (BMPs), collagen I, and Runx2. Cells were also capable of depositing a mineralized matrix, confirming the commitment of these cells to the osteogenic lineage. When FGFR1 activity was blocked, a reduction in cell proliferation and a coincident upregulation of Runx2 with enhanced mineralization of cultures was observed. These results indicate that FGFRs play critical roles in cell recruitment and differentiation during the process of osteogenesis in mES cells. In particular, the data indicate that FGFR1 plays a pivotal role in osteoblast lineage determination.
authors.affiliation: Stem Cell and Tissue Repair Laboratory, Institute of Molecular and Cell Biology, Proteos, Singapore 138673.
authors.forename: Kee
authors.initials: K
authors.lastname: Woei Ng
authors.forename: Tobias
authors.initials: T
authors.lastname: Speicher
authors.forename: Christian
authors.initials: C
authors.lastname: Dombrowski
authors.forename: Torben
authors.initials: T
authors.lastname: Helledie
authors.forename: Larisa M
authors.initials: LM
authors.lastname: Haupt
authors.forename: Victor
authors.initials: V
authors.lastname: Nurcombe
authors.forename: Simon M
authors.initials: SM
authors.lastname: Cool
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage; D048430:Cell Shape; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D010012:Osteogenesis / Q000502:physiology*; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
chemical_list: D015415:Biomarkers; D017468:Receptors, Fibroblast Growth Factor