Development of technical bases for biotooth formation by using iPS cells

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Induced pluripotent stem(iPS) cells are generated from somatic cells by the simultaneous introduction of several factors ; and they differentiate into the3embryonic germlayers with an extensive proliferative capacity. Recently, many researchers have reported that iPS cells can differentiate into different cell types, such as neurons, cardiac myocytes, and renal lineage cells, under appropriate conditions. Therefore, iPS cells have emerged as potential cell sources for tooth regeneration. In this study, we aimed to form biotooth by recombination of iPS cell-induced dental germ cells with mouse dental germ cells. First, we established the induction methods of iPS into mesenchymal stem cell(MSC). The differentiated cells expressed STRO-1, MSC marker, more than90%, as well as other MSC markers. Further, we established a culture protocol to induce the differentiation of iPS cells into neural crest like cells(NCLC), which is a precursor cell of dental mesenchymal cells. After the induction, the most of cells expressed neural crest cells markers. The cells did not form teratomas after transplantation into nude mice subcutaneously, suggesting the safety for clinical applications. Moreover, in recombination cultures of NCLC and mousedental epithelium, NCLC exhibited a gene expression pattern involving dental mesenchymal cells. Some NCLC also expressed dentin sialoprotein. Conditioned medium of mouse dental epithelium cultures further enhanced the differentiation of NCLC into odontoblasts. These results suggest that iPS cells are useful cell sources for tooth regeneration and tooth development studies.