Functional Tooth Regeneration.
Three-dimensional organogenesis in vivo is principally regulated by the spatiotemporal developmental process that relies on the cellular behavior such as cell growth, migration, differentiation, and cell-to-cell interaction. Organ development and morphogenesis have been elucidated to be regulated by the proper transient expression of various signaling molecules including cytokines, extracellular matrix, and adhesion molecules based on the epithelial and mesenchymal interactions. Current bioengineering technology for regenerating three-dimensional organ has progressed to the replication of organogenesis, thereby enabling the development of fully functional bioengineered organs using bioengineered organ germs that are generated from immature stem cells via tissue engineering technology in vitro.To achieve precise replication of organogenesis, we have developed a novel three-dimensional cell manipulation method designated the organ germ method, and enabled the generation of a structurally correct and fully functional bioengineered tooth in vivo. This method is also expected to be utilized for analyzing gene and protein functions during organogenesis. Here, we describe protocols for the tooth germ reconstitution by using the organ germ method and for the functional analysis of tooth development in vitro and in vivo.