title: Mouse ES cells have a potential to differentiate into odontoblast-like cells using hanging drop method.
abstract: OBJECTIVE
  We examined whether mouse embryonic stem (ES) cells can differentiate into odontoblast-like cells without epithelial-mesenchymal interaction.
  
  
  METHODS
  Cells were cultured by the 'hanging drop' method using a collagen type-I scaffold (CS) combined with bone morphogenetic protein (BMP)-4 (CS/BMP-4). Expression of odontoblast-related mRNA and protein, and cell proliferation were performed by reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence staining and WST-1 assay, respectively.
  
  
  RESULTS
  Cells potently expressed odontoblast-related cell marker mRNAs following induction of odontoblastic differentiation. Dentin sialophosphoprotein, a marker of mature odontoblasts, was strongly expressed in differentiated ES cells. The cells also acquired an odontoblast-like functional phenotype, as evidenced by the appearance of alkaline phosphatase activity and calcification. The cell-surface expression of α2, α6, αV and αVβ3 integrin proteins was rapidly upregulated in differentiated cells. Finally, anti-α2 integrin antibody suppressed the expression of odontoblastic markers in cells grown using this culture system, suggesting that α2 integrin expression in ES cells triggers their differentiation into odontoblast-like cells.
  
  
  CONCLUSIONS
  Mouse ES cells cultured by the 'hanging drop' method are able to differentiate into cells with odontoblast-specific physiological functions and cell-surface integrin protein expression.
authors.affiliation: Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Kawai
authors.forename: N
authors.initials: N
authors.lastname: Ozeki
authors.forename: H
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: T
authors.initials: T
authors.lastname: Tanaka
authors.forename: K
authors.initials: K
authors.lastname: Nakata
authors.forename: M
authors.initials: M
authors.lastname: Mogi
authors.forename: H
authors.initials: H
authors.lastname: Nakamura
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology*; D051379:Mice; D009804:Odontoblasts / Q000166:cytology*
keywords: cell differentiation; cell-matrix interactions; collagen; integrin; odontoblast; stem cell