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. 2024 Aug 7;10(15):e35295.
doi: 10.1016/j.heliyon.2024.e35295. eCollection 2024 Aug 15.

Differentiation of cultured hair follicle neural crest stem cells into functional melanocytes

Hongjuan Wang  1   2   3 , Wen Hu  1   2   3 , Fang Xiang  1   2   3 , Zixian Lei  1   2   3 , Xiangyue Zhang  1   2   3 , Jingzhan Zhang  1   2   3 , Yuan Ding  1   2   3 , Xiaojing Kang  1   2   3
Affiliations

Differentiation of cultured hair follicle neural crest stem cells into functional melanocytes

Hongjuan Wang et al. Heliyon. .

Abstract

Many autologous melanocytes are required for surgical treatment of depigmentation diseases such as vitiligo. However, primary cultured melanocytes have a limited number of in vitro passages. The production of functional epidermal melanocytes from stem cells provides an unprecedented source of cell therapy for vitiligo. This study explores the clinical application of melanocytes induced by hair follicle neural crest stem cells (HFNCSCs). This study established an in vitro differentiation model of HFNCSCs into melanocytes. Results demonstrate that most differentiated melanocytes expressed the proteins C-KIT, MITF, S-100B, TYRP1, TYRP2, and tyrosinase. The HFNCSC-derived melanocytes were successfully transplanted onto the dorsal skin of mice and survived in the local tissues, expressing marker protein of melanocytes. In conclusion, HFNCSCs in mice can be induced to differentiate into melanocytes under specific conditions. These induced melanocytes exhibit the potential to facilitate repigmentation in the lesion areas of vitiligo-affected mice, suggesting a promising avenue for therapeutic intervention.

Keywords: Differentiation; Hair follicle neural crest stem cells (HFNCSCs); Melanocyte; Transplantation; Vitiligo.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
Primary culture HFNCSCs and immunofluorescence results of HFNCSCs (scale bar = 100 μm). (a) A small population of HFNCSCs emerging from tissue blocks. (b) After 12–14 days, HFNCSC achieved a confluence of 60 %–70 %; (c) Approximately 20 days later, HFNCSC reached a confluence of 80 %–90 %, mostly exhibiting spindle-shaped and triangular morphologies. (d) The HFNCSCs showed positive staining for nestin and SOX10.
Fig. 2
Fig. 2
Characterization of HFNCSCs-derived melanocytes(scale bar = 100 μm). (a)Microscopy images of HFNCSCs. (b) Microscopy images of melanocytic differentiation from HFNCSC. (c) l-DOPA staining: The cytoplasm and dendrites of the cells appeared in black or dark, indicating a positive reaction. (d) Immunofluorescence staining of C-KIT, MITf, S–100B, TYRP1, TYRP2, and tyrosinase in melanocytes (scale bar = 100 μm). (e) The fluorescence expression of C-KIT, TYRP2, tyrosinase, MITF, S-100, and TYRP1 in melanocytes (n = 3),*P < 0.05, ***P < 0.001.
Fig. 3
Fig. 3
Simultaneous grafting of melanocytes in mice. (a) On the 36th day, evident depigmentation was observed in the modeling area of mice in the Model group and Transplantation group. (b) By the 52nd day, significant pigmentation was noticeable in the Transplantation group. (c) In comparison to the Model group, HE staining in the Transplantation group revealed a thinner epidermis at the lesion site, reduced inflammation infiltration, and an increased number of hair follicles.
Fig. 4
Fig. 4
Expression of antibodies in skin tissues. (a) Expression of c-KIT, MITF, S100 B, tyrosinase, TYRP1, and TYRP2 in skin tissues. (b) The fluorescence was quantified using ImageJ (n = 3). **: P < 0.01 vs Control, ***: P < 0.001 vs Control; ##: P < 0.01 vs Model, ###: P < 0.001 vs Model.
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