Human fibroblasts treated with hydrogen peroxide stimulate human melanoblast proliferation and melanocyte differentiation, but inhibit melanocyte proliferation in serum-free co-culture system.
BACKGROUND
Oxidative stress caused by hydrogen peroxide (H2O2) elicits harmful effects on human melanocytes such as DNA damage and cell death. On the contrary, H2O2is known to possess beneficial effects on melanocytes. However, mechanisms of the beneficial effects of H2O2on melanocytes have not been fully understood, especially the indirect effects on melanocyte proliferation and differentiation from cells constituting surrounding tissue environment such as fibroblasts.


OBJECTIVE
The aim of this study was to clarify whether H2O2-treated human fibroblasts affect the proliferation and differentiation of human melanocytes using serum-free co-culture system.


METHODS
Epidermal melanoblasts and melanocytes were co-cultured with H2O2-treated or control fibroblasts in serum-free culture media. The effects of H2O2-treated fibroblasts were detected by changes in the proliferation and differentiation of melanoblasts/melanocytes.


RESULTS
H2O2-treated fibroblasts stimulated the proliferation of melanoblasts and the differentiation, melanogenesis, and dendritogenesis of melanocytes, but inhibited the proliferation of melanocytes. In the melanocytes co-cultured with H2O2-treated fibroblasts, the expression of tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1), and KIT was increased, whereas TYRP2 and microphthalmia-associated transcription factor showed no change.


CONCLUSIONS
These results suggest that H2O2-treated fibroblasts can produce and release some mitogenic and melanogenic factors toward melanoblasts in addition to some proliferation-inhibiting factors toward melanocytes. The stimulation of melanocyte differentiation seems to be performed through the upregulation of TYR, TYRP1, and KIT.
Fukushima Project Headquarters, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555, Japan. Electronic address: hirobe.tomohisa@qst.go.jp.
Tomohisa
T
Hirobe
Shiseido Global Innovation Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama 224-8558, Japan.
Tatako
T
Shibata
Shiseido Global Innovation Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama 224-8558, Japan.
Kiyoshi
K
Sato
D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods*; D016895:Culture Media, Serum-Free / Q000737:chemistry; D004249:DNA Damage; D004817:Epidermis / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry*; D008543:Melanins / Q000378:metabolism; D008544:Melanocytes / Q000166:cytology*; D051739:Microphthalmia-Associated Transcription Factor / Q000378:metabolism; D008856:Microscopy, Fluorescence; D014442:Monophenol Monooxygenase / Q000378:metabolism; D018384:Oxidative Stress; D010858:Pigmentation
D016895:Culture Media, Serum-Free; D008543:Melanins; D051739:Microphthalmia-Associated Transcription Factor; D006861:Hydrogen Peroxide; D014442:Monophenol Monooxygenase
Differentiation; Fibroblast; Hydrogen peroxide; Melanocyte; Proliferation