| 研究実績の概要 |
The purpose of this study was to identify new methods and culture conditions to enhance chondrogenic differentiation of human bone marrow-derived mesenchymal stem/progenitor cells (hBMSCs) by applying chemical, physical and biological cues, including overexpression of DNA methyltransferases (DNMTs) in cells. We attempted to fabricate a chemically- and physically-controlled microenvironment using materials (e.g., hydrogels) that recapitulate native cell niche characteristics to optimally modulate the chondrogenic differentiation of BMSCs. Besides the physical and chemical stimulations, we also evaluated the chondrogenic differentiation of hBMSCs after overexpression of DNMTs that eventually induce DNA methylation at CPG rich regions of key genes. To study the effect of DNMTs on chondrogenesis of hBMSCs, we transfected overexpression vectors of DNMT3A, DNMT3B and the pcDNA control in hBMSCs, and culture the cells in a 3D micromass culture system with growth factors and glucocorticoids.
Additionally, since in vitro synthesized cartilage tissue generally undergoes through mineralization, we also attempted to understand the mechanisms involved in the chondrocyte death associated with initial mineralization process by using the secondary ossification center of mouse femur as in vivo model.
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