Fig. 1

hESC-derived pigmented cells exhibit a melanocytic phenotype. A Graphical illustration of differentiating from embryonic stem cells to melanocytes. B Pigmented and dendritic cells appeared 7–10 days after exposure to puromycin at passage 0 (p0). C Pigmented cells increased, while non-pigmented cells decreased around passage 9 (p9). D Pigmented cells were dominant by passage 15. E Gross appearance of pigmented cells. The culture plate was covered with black colonies (left), and the cell pellet was black without any staining (right). F, G Human Embryonic stem cells-derived melanocytes (F: EMCs) at passage 25 resembled tissue-separated melanocytes (G: TMCs) in morphology. H Quantitative RT-PCR analysis of the genes for NANOG, OCT4, TYRP1, MITF, Tyrosinase, PAX3, Kit, and DCT, in undifferentiated cells (UDCs), EMCs, and TMCs. The expression levels were normalized by expressions of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The expression levels of TMC were set at 1.0. Each expression level was calculated from the results of triplicate technical experiments and the charts are drawn as the average ± standard deviation. I Immunocytochemical analysis of EMCs(left) and TMCs (right) with antibodies against MITF (red), Tyrosinase (red), and MelanA (red). Nuclei were counterstained with DAPI (blue)