Abstract
Hair follicles harbor a heterogeneous regenerative cell pool and represent a putative low-to-non-invasively available source of stem cells. We previously reported a technology for culturing human melanocytes from the hair follicle outer root sheath (ORS) for autologous pigmentation of tissue engineered skin equivalents. This study translated the ORS technology to horses. We de-veloped a culture of equine melanocytes from the ORS (eMORS) from equine forelock hair follicles cultured by means of an analogue human hair follicle-based in vitro methodology. The procedure was adjusted to equine physiology by addition of equine serum to the culture medium. The hair follicles were isolated by macerating forelock skin rests, enzymatically digested and subjected to air-medium-interface cultivation method. The procedure resulted in differentiated equine melanocytes, which exhibited typical morphology, presence of melanosomes, expression of cytoskeleton proteins vimentin, α-SMA, Sox2, S100ß and tyrosinase as well as tyrosinase activity followed by production of melanin. According to all assessed parameters, eMORS could be ranked as partially melanotic melanocytes. The results of the study offer an experimental base for further insight into hair follicle biology in equine and for comparative studies of hair follicles across different species.
Highlights
The therapeutic potential of adult stem cells has putatively fueled regenerative medicine research in the several past decades
Stem cells residing in the outer root sheath (ORS) of the hair follicle were identified in the bulge area adjacent to the sebaceous gland [1]
This study presents a re-visited method for obtaining HUMORS applied to horse forelock hair follicles in order to culture equine melanocytes from the outer root sheath
Summary
The therapeutic potential of adult stem cells has putatively fueled regenerative medicine research in the several past decades. The standard methods of obtaining adult stem cells have been relying on invasive, biopsy-based approaches, which involved substantial donor site morbidity. The alternative, less invasive methods of harvesting stem cells have been progressively gaining momentum. To this day, the hair follicle has remained the least invasively obtainable source of stem cells as well as the one with the most favorable sample/yield ratio. ORS stem cell pluripotency and their potential in regenerative medicine were shown in subsequent studies [2,3].
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