Craniofacial Bone Regeneration using iPS Cell-Derived Neural Crest Like Cells

Abstract

Induced pluripotent stem (iPS) cells represent a powerful source for cell-based tissue regeneration because they are patient-specific cells and can differentiate into specialized cell types. Previously, we have demonstrated the derivation of neural crest like cells from iPS cells (iPS-NCLCs), and these cells have the potential to differentiate into dental mesenchymal cells, which subsequently differentiate into odontoblasts and dental pulp cells. In this study, we show that iPS-NCLCs can differentiate into mesenchymal stem cells (iPS-NCLC-MSCs), which contribute to craniofacial bone regeneration. iPS-NCLCs were cultured in serum-containing media and differentiated into functional MSCs, as confirmed by expression MSC markers and their ability to differentiate into osteoblasts, adipocytes, and chondrocytes in vitro. iPS-NCLC-MSCs were negative for markers of undifferentiated iPS cells and did not develop into teratomas when transplanted to immunodeficient mice. Further, iPS-NCLC-MSCs grew normally and differentiated into osteoblasts on hydroxyapatite scaffolds in vitro. To assess the potential of iPS-NCLC-MSCs to regenerate craniofacial bone in vivo, iPS-NCLC-MSCs were transplanted into critical-size calvarial defects in immunodeficient mice for 8 weeks. Histological analysis revealed that iPS-NCLC-MSCs differentiated into osteoblasts and contributed to bone regeneration without tumor formation. These results indicate that iPS-NCLC-MSCs could be a potential candidate for cell-based craniofacial bone tissue repair and regeneration.