Melanocyte Chitosan/Gelatin Composite Fabrication with Human Outer Root Sheath-Derived Cells to Produce Pigment

Chuan Gu,Fei Liu,Huitang Xia,Jinny Huang,Guangdong Zhou,Xianyu Zhou,Xusong Luo,Xiuxia Wang,Yan Ma,Jun Yang

doi:10.1038/s41598-019-41611-5

Chuan Gu, Fei Liu + Show 8 more

Open Access

https://doi.org/10.1038/s41598-019-41611-5

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Abstract

The hair follicle serves as a melanocyte reservoir for both hair and skin pigmentation. Melanocyte stem cells (MelSCs) and melanocyte progenitors reside in the bulge/sub-bulge region of the lower permanent portion of the hair follicle and play a vital role for repigmentation in vitiligo. It would be beneficial to isolate MelSCs in order to further study their function in pigmentary disorders; however, due to the lack of specific molecular surface markers, this has not yet been successfully accomplished in human hair follicles (HuHF). One potential method for MelSCs isolation is the “side population” technique, which is frequently used to isolate hematopoietic and tumor stem cells. In the present study, we decided to isolate HuHF MelSCs using “side population” to investigate their melanotic function. By analyzing mRNA expression of TYR, SOX10, and MITF, melanosome structure, and immunofluorescence with melanocyte-specific markers, we revealed that the SP-fraction contained MelSCs with an admixture of differentiated melanocytes. Furthermore, our in vivo studies indicated that differentiated SP-fraction cells, when fabricated into a cell-chitosan/gelatin composite, could transiently repopulate immunologically compromised mice skin to regain pigmentation. In summary, the SP technique is capable of isolating HuHF MelSCs that can potentially be used to repopulate skin for pigmentation.

Highlights

Www.nature.com/scientificreports sign of Melanocyte stem cells (MelSCs) depletion in the hair follcile[11,12]
Www.nature.com/scientificreports sign of MelSCs depletion in the hair follcile[11,12]. These findings indicate that MelSCs are of high importance and are the real melanocyte reservoir for repigmentation in the hair shaft and skin epidermis[13]
This assay is based on the capability of side population” (SP) cells to better efflux the DNA-binding dye, Hoechst 3334228, or mitochondrial-binding dye, Rhodamine12332 because they have a higher number of ATP-binding cassette (ABC) transporter proteins expressed within the cell matrix

Summary

Introduction

Www.nature.com/scientificreports sign of MelSCs depletion in the hair follcile[11,12]. Methods like NCORSHFS that involve repigmentation through cellular grafting are generally used for small depigmentary lesions They are not appropriate for more massive lesions due to the lack of sufficient donor hair sources, especially if the patient suffers from hair loss caused by other diseases such as alopecia areta or scalp burn scar. These disadvantages impede the broader application of NCORSHFS and prompt us to exploit a more economical way to efficiently obtain favorable MCs from a small number of hair follicles to treat vitiligo, such as in vitro isolation of MelSCs and cell propagation with extended cell culture. This could potentially help broaden the application of using MelSCs and cultured follicular melanocytes to treat skin pigmentary disorders

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