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. 2022 Apr 19;12:6416. doi: 10.1038/s41598-022-08163-7

Figure 1.

Figure 1

Differentiation of human induced pluripotent stem cell-derived melanocytes. (a) Schematic diagram of melanocyte differentiation from hiPSCs. (b) Morphology of cells at each stage of differentiation. Scale bar indicates 100 µm. (c) Image of day 28 cell pellets showing a darkly pigmented phenotype (left). Pigmented cells visualized under a bright-field microscope (right). Arrowheads indicate pigmented cells. Scale bar indicates 100 µm. (d) Transmission electron microscopy (TEM) image of melanosomes in hiMels. Scale bar indicates 2 µm. (e) Gene expression of the pluripotency marker OCT4 and melanocyte markers PAX3, SOX10, MITF, TYRP1, and TYR. The relative gene expression values were calculated using the delta cycle-threshold (ΔCt) method. (f) Bright-field microscope images of hiPSCs, hiMels, and NHEMs. (g) L-DOPA staining of hiMels. Scale bar indicates 100 µm. (h) Fontana–Masson staining image of hiMels. (i) Immunocytochemical analysis of melanocyte marker TRP1 (red) and DAPI (blue). Scale bar indicates 100 µm. (j) Quantification of tyrosinase activity in hiMels. Data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 indicate statistical significance. ns non-significant, hiPSCs human induced pluripotent stem cells, hiMels human hiPSCs derived melanocytes, L-DOPA 3,4-dihydroxyphenylalanine, NHEM normal human epidermal melanocyte.