Embryonic development relies on precise execution of differentiation programs carried out by pluripotent stem cells that have the ability to form every cell in the body. During gastrulation, these cells become further specified into endodermal, mesodermal, ectodermal, and neural crest derivations. Dental-oral-craniofacial (DOC) tissues such as teeth, facial bones and salivary glands are generated by the neural crest in a process that often requires input from adjacent non-neural ectoderm, and bones of the cranial vault and facial muscles derive from mesoderm. However, the molecular mechanisms underlying the formation of DOC tissues have remained less well understood. This symposium will describe recent advances in our understanding of differentiation into DOC tissues by highlighting diverse approaches and novel technologies. In particular, through proteomics and ATAC seq, new insight is emerging on the mechanisms by which pluripotent stem cells commit to a neural crest fate and the role of ubiquitylation in this process. Novel live cell imaging and spatial genomic analysis is shedding light on how neural crest cells gain and maintain their stemness. By using single-cell analysis, lineage trajectories of stem and progenitor cells are being used to map out development of salivary glands. Furthermore, directed differentiation following embryonic pathways of specification is being applied to generate neural crest-specific and mesoderm-specific osteoprogenitor populations with signature transcriptomic patterns. These studies provide the opportunity to better identify and characterize DOC stems cells, to determine the pathogenetic roles that they play in DOC diseases, and to use them to restore normal structure and function.