Methods for Derivation of Multipotent Neural Crest Cells Derived from Human Pluripotent Stem Cells.

Multipotent, neural crest cells (NCCs) produce a wide range of cell types during embryonic development. This includes melanocytes, peripheral neurons, smooth muscle cells, osteocytes, chondrocytes, and adipocytes. The protocol described here allows for highly efficient differentiation of human pluripotent stem cells to a neural crest fate within 15 days. This is accomplished under feeder-free conditions, using chemically defined medium supplemented with two small molecule inhibitors that block glycogen synthase kinase 3 (GSK3) and bone morphogenic protein (BMP) signaling. This technology is well suited as a platform to understand in greater detail the pathogenesis of human disease associated with impaired neural crest development/migration.