Saaya Koike

A quantitative analysis of multilineage-differentiating stress-enduring (Muse) cells in human adipose tissue and efficacy of melanocytes induction

We have shown previously that multilineage-differentiating stress-enduring (Muse) cells in neonatal fibroblasts can differentiate into functional melanocytes. In this study, we quantitate Muse cells in adipose-mesenchymal stem cells (adipose-MSCs) of human subcutaneous tissue obtained from 11 subjects of various ages, and measured efficacy of melanocytes induction from Adipose-MSC-derived Muse cells (hASC-Muse cells). There was a statistically significant negative correlation between the age of donors and the numbers of adipose-MSCs recovered per g fat as well as the percentage of SSEA3+ cells in the adipose-MSC populations, but isolated hASC-Muse cells showed pluripotency and growth curves equally regardless the age of donors. Adipose-Muse cells sequentially expressed melanocyte-related genes including KIT, MITF, TYRP1 PMEL, DCT, melanocortin 1 receptor (MC1R), and TYR at a comparable level to melanocytes during 6-week culture. Parallel with MC1R expression, adipose-Muse cells increased melanin content by α-MSH stimulation. By quantitating the cell numbers recovered at each step, we found that 10g of adipose tissue could produce at least 2.5×106 melanocytes after 6 weeks of culture. These studies suggest that induction of melanocytes from adipose-Muse is a novel approach to obtain sufficient numbers of melanocytes for clinical application and in vitro study of melanocyte differentiation.

Toll-like receptors 2 and 3 enhance melanogenesis and melanosome transport in human melanocytes

Because little is known about how the innate immune response influences skin pigmentation, we examined whether Toll‐like receptor (TLR) agonists participate in melanogenesis and melanosome transportation. We observed that TLR2/2 agonist HKLM and TLR3 agonist Poly(I:C) increased the amount of extracellular melanin from primary human epidermal melanocytes. HKLM, but not Poly(I:C), increased the melanogenic genes such as tyrosinase and dopachrome tautomerase. Poly(I:C) increased the expression of Rab27A, a molecule that facilitates melanosome transport to perimembranous actin filament. UVB irradiation induced Rab27A and melanosome transportation in a similar manner of Poly(I:C). SiRNA for TLR3 or Rab27A suppressed the perimembranous accumulation of Gp100‐positive vesicles in melanocytes and decreased melanin transfer to neighboring keratinocytes induced by both Poly(I:C) and UVB. These results suggest that the microenvironment in the epidermis and innate immune stimuli, such as microbiome and ultraviolet represented here by TLR2 and TLR3 agonists, could affect the melanogenesis in human melanocytes.