RRC ID 44464
Author Kawai R, Ozeki N, Yamaguchi H, Tanaka T, Nakata K, Mogi M, Nakamura H.
Title Mouse ES cells have a potential to differentiate into odontoblast-like cells using hanging drop method.
Journal Oral Dis
Abstract 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.
Volume 20(4)
Pages 395-403
Published 2014-5-1
DOI 10.1111/odi.12134
PMID 23731055
MeSH Animals Cell Differentiation* Cells, Cultured Cytological Techniques / methods Embryonic Stem Cells / cytology* Mice Odontoblasts / cytology*
IF 2.613
Times Cited 15
WOS Category DENTISTRY, ORAL SURGERY & MEDICINE
Resource
Human and Animal Cells B6G-2(AES0003)