The effects of NB-UVB on the hair follicle-derived neural crest stem cells differentiating into melanocyte lineage in vitro

Abstract

Narrow-band UVB (NB-UVB) is an effective therapeutic option in the treatment of vitiligo. Despite the apparent clinical efficacy, the underlying mechanism of how topical NB-UVB induces repigmentation in vitiligo has not been clearly elucidated. To investigate the effects of NB-UVB on the maturation of melanocyte lineage differentiated from hair follicle-derived neural crest stem cells (HF-NCSCs) in vitro. HF-NCSCs were isolated from mouse whisker follicles. The isolated cells were multipotent and expressed embryonic NCSC biomarkers. The effects of NB-UVB on development and differentiation of HF-NCSCs were evaluated. We assessed cell viability, melanogenesis and migration of melanocytes derived from HF-NCSCs after NB-UVB radiation. Tyrosinase, Tyrp1, Dct, Kit, Mc1R, Fzd4, NT3R, Ednra, EP1, TGFβR, Sox10, Mitf, Lef1 and Pax3 gene expression was measured by quantitative RT-PCR, while Tyrosinase, Sox10 and Mitf protein expression were measured by Western blot analysis. Cell migration was measured by Boyden chamber transwell assay. NB-UVB increased the expression of tyrosinase during melanocytic differentiation from mouse HF-NCSCs, however, NB-UVB inhibited proliferation of melanocytes derived from HF-NCSCs. Mechanistically, increased melanocyte maturation after NB-UVB treatment was resulted from increased expression of several key melanogenic factors, including Sox10, Kit and Mc1R, which play a critical role to promote tyrosinase expression. Furthermore, the migration of the HF-NCSCs-derived melanocytes was downregulated as NB-UVB doses increased. However, the migration of HF-NCSCs was upregulated under 0.4J NB-UVB radiation. Those data provide in vitro evidence demonstrating some direct effects of NB-UVB on pigmentation of melanocyte lineage differentiated from HF-NCSCs, and may provide a possible mechanism for the effect of NB-UVB in vitiligo.