title: Feeder-free Derivation of Melanocytes from Human Pluripotent Stem Cells.
abstract: Human pluripotent stem cells (hPSCs) represent a platform to study human development in vitro under both normal and disease conditions. Researchers can direct the differentiation of hPSCs into the cell type of interest by manipulating the culture conditions to recapitulate signals seen during development. One such cell type is the melanocyte, a pigment-producing cell of neural crest (NC) origin responsible for protecting the skin against UV irradiation. This protocol presents an extension of a currently available in vitro Neural Crest differentiation protocol from hPSCs to further differentiate NC into fully pigmented melanocytes. Melanocyte precursors can be enriched from the Neural Crest protocol via a timed exposure to activators of WNT, BMP, and EDN3 signaling under dual-SMAD-inhibition conditions. The resultant melanocyte precursors are then purified and matured into fully pigmented melanocytes by culture in a selective medium. The resultant melanocytes are fully pigmented and stain appropriately for proteins characteristic of mature melanocytes.
authors.affiliation: The Center for Stem Cell Biology, Developmental Biology Program, Memorial Sloan-Kettering Cancer Center; Cancer Biology and Genetics Program, Gerstner Sloan-Kettering Graduate School, Sloan-Kettering Institute for Cancer Research.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Callahan
authors.affiliation: Thermo Fisher Scientific.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Mica
authors.affiliation: The Center for Stem Cell Biology, Developmental Biology Program, Memorial Sloan-Kettering Cancer Center; studerl@MSKCC.ORG.
authors.forename: Lorenz
authors.initials: L
authors.lastname: Studer
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D009432:Neural Crest / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*