Improved method of differentiation, selection and amplification of human melanocytes from the hair follicle cell pool

Abstract

Hair root harbours a complex cell pool with an immense developmental potential. Several lineages, including skin, can be differentiated from the multipotent to pluripotent cells of outer root sheath (ORS) of hair follicle. Outer root sheath presents the most opulent non-invasively gained adult stem cell source known. For the purposes of cultivating melanocytes designated for graft-based treatments of depigmentation disorders, we have established an ex vivo/in vitro cultivation method by introducing several methodological improvements to the ORS explant method of Dieckmann. As a result, we gained a higher, purer yield of differentiated melanocytes in half the time (at least 10 6 of 95% pure cells in 4 weeks). This reliable cultivation procedure begins with the epilation of 60 hairs and yields high numbers of ORS melanocytes that could be used for grafting applications. The procedure not only utilises the developmental potential of hair root cell pool and favors differentiation into melanocytes, but also contributes to the general trend of minimal-to-non-invasive strategies for regenerative medicine.

Sulfated hyaluronan‐containing artificial extracellular matrices promote proliferation of keratinocytes and melanotic phenotype of melanocytes from the outer root sheath of hair follicles

“…1) were synthesized and characterized as reported previously. 13 C NMR spectra were recorded in D 2 O with a 400 MHz device (Bruker Corporation, 5 mm BBFO smart probe) in order to analyze the substituent distribution of sHA derivatives. The following sulfation reaction of TBA-HA was performed under argon in DMF at room temperature.…”

“…13 C NMR spectra of hyaluronic acid and of the sulfated derivatives sHA1 and sHA4. quantification was calculated by the means of the ΔΔC T method 64 following calibration to the value of PS cultures and set at 1.0.…”

Melanocyte stem cells as potential therapeutics in skin disorders

“…As ORS cells include pluripotent neural crest stem cell (NCSC)-like stem cells and quiescent stem cells that have potential to be differentiated into various cell types, ORS cells are a valuable resource in regenerative medicine [106] . Dieckmann et al demonstrated methods to isolate melanocytes from ORS cells in human anagen hair and propagated them in vivo; moreover, the yields of the melanocytes could be improved by a sequential method [107,108] . Although this method is not yet used in the clinical setting, the source of the melanocytes could be the MelSCs in ORS and this represents a promising method for biologic therapeutics in the near future.…”

Polycaprolactone fiber meshes provide a 3D environment suitable for cultivation and differentiation of melanocytes from the outer root sheath of hair follicle

“…Most of the ORS subpopulations are promising candidates for regenerative therapies. The ORS cell pool is easily accessible by harmless plucking of hair and therefore valuable for approaches that require non‐invasive sampling, whether for research or therapeutic purposes …”