Abstract
Recently, multipotent stem cells have been studied as material for regeneration of missing craniofacial bone. Almost all craniofacial structures are derived from cranial neural crest cells. We investigated the possibility of craniofacial bone induction from mouse embryonic stem (ES) cells differentiated into neural crest via neuroectoderm.
First, we cultured mouse ES cells in serum-free neuroinduction medium for sphere formation and confirmed the existence of precursors of neuronal and neural crest cells by immunostaining using specific antibodies for Nestin and AP2α, respectively. The sphere contained Nestin and AP2α positive cells, indicating that a portion of the ES cells differentiated into neural crest cells through neural precursor cells during sphere formation. Subsequently, these spheres were transferred to an alternative plate and grown for 18 days in osteogenic medium containing dexamethasone. Alizarin red S staining revealed mineral deposition, suggesting that the cells within the spheres were induced to osteogenesis. These findings indicated the possibility that craniofacial bone can be generated from mouse ES cells through spheres cultured in serum-free medium.
