Mouse ES cells have a potential to differentiate into odontoblast-like cells using hanging drop method.
OBJECTIVES
We examined whether mouse embryonic stem (ES) cells can differentiate into odontoblast-like cells without epithelial-mesenchymal interaction.


MATERIALS AND 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.
['Kawai|R|R|', 'Ozeki|N|N|', 'Yamaguchi|H|H|', 'Tanaka|T|T|', 'Nakata|K|K|', 'Mogi|M|M|', 'Nakamura|H|H|']
D000818:Animals D002454:Cell Differentiation D002478:Cells, Cultured D003584:Cytological Techniques D053595:Embryonic Stem Cells D051379:Mice D009804:Odontoblasts
collagen odontoblast integrin cell differentiation cell-matrix interactions stem cell