title: Generation of melanocytes from induced pluripotent stem cells.
abstract: Epidermal melanocytes have an important role in protecting skin from UV rays, and are implicated in a variety of skin diseases. Here, we developed an efficient method for differentiating induced pluripotent stem cells (iPSCs) into melanocytes. We first generated iPSCs from adult mouse tail-tip fibroblasts (TTFs) using retroviral vectors or virus-free piggyBac transposon vectors carrying murine Sox2, Oct3/4, c-Myc, and Klf4. The TTF-derived iPSC clones exhibited similar morphology and growth properties as mouse embryonic stem (ES) cells. The iPSCs expressed ES cell markers, displayed characteristic epigenetic changes, and formed teratomas with all three germ layers. The iPSCs were used to generate embryonic bodies and were then successfully differentiated into melanocytes by treatment with growth factors. The iPSC-derived melanocytes expressed characteristic melanocyte markers and produced melanin pigment. Electron microscopy showed that the melanocytes contained mature melanosomes. We manipulated the conditions used to differentiate iPSCs to melanocytes and discovered that Wnt3a is not required for mouse melanocyte differentiation. This report shows that melanocytes can be readily generated from iPSCs, providing a powerful resource for the in vitro study of melanocyte developmental biology and diseases. By inducing iPSCs without viruses, the possibility of integration mutagenesis is alleviated, and these iPSCs are more compatible for cell replacement therapies.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Ruifeng
authors.initials: R
authors.lastname: Yang
authors.forename: Min
authors.initials: M
authors.lastname: Jiang
authors.forename: Suresh M
authors.initials: SM
authors.lastname: Kumar
authors.forename: Theodore
authors.initials: T
authors.lastname: Xu
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Leihong
authors.initials: L
authors.lastname: Xiang
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Xu
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D058732:Embryoid Bodies / Q000187:drug effects; D053595:Embryonic Stem Cells; D005347:Fibroblasts / Q000187:drug effects; D016259:Genes, myc; D005822:Genetic Vectors; D005855:Germ Layers / Q000187:drug effects / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D008543:Melanins / Q000096:biosynthesis; D008544:Melanocytes / Q000166:cytology*; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics; D013724:Teratoma / Q000235:genetics; D023822:Tissue Engineering / Q000379:methods*; D060509:Wnt3A Protein / Q000378:metabolism
chemical_list: D015415:Biomarkers; D004251:DNA Transposable Elements; D036341:Intercellular Signaling Peptides and Proteins; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D008543:Melanins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse