Induction of Mandibular bone and tooth germ in embryo body formation from mouse and human iPS cells in serum-free cell culture

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The miPS cells have been maintained in ESF7 serum-free medium for more than 2 years with an undifferentiated phenotype. An FGF-2 induced miPS cell differentiation into tooth germ progenitor cells. hiPSCs are commonly maintained on feeder cells in medium supplemented with FBS. In this study, we successfully generated hiPSCs from human dental pulp cells using Oct3/4, Sox2, Klf4, c-Myc with retroviral vectors in serum- and feeder-free defined culture conditions. These hiPSC retained the property of selfrenewal, and pluripotency evaluated by differentiation into derivatives of all three primary germ layers by embryo body formation assay and teratoma formation assay. hiPSCs cultured in hESF9 medium absolutely required TGF-beta1 to maintain pluripotency. This simple serum-free adherent monoculture system will allow us to elucidate the cell responses to growth factors under defined conditions and can eliminate the risk might be brought by undefined pathogens.