Abstract
Facial epidermal pigmentation and skin tumors can be caused by UV exposure and other physical and chemical irritations. In this report we describe the primary culture of melanocytes from human face skin. The ability to culture these melanocytes will enable their morphological and biological properties to be investigated. Skin specimens were obtained from patients who had undergone lower blepharoplasty procedures. Digestion with neutral protease and trypsin was used to obtain single cell suspensions of epidermal cells. The cells were cultured in M254 medium supplemented with human melanocyte growth solution. Cell morphology was observed using inverted microscopy. Melanocytes were positively identified using both l-DOPA staining and S-100 protein immunohistochemical staining. Immunofluorescence was used to confirm the expression of tyrosinase-related protein-1, a melanocyte-specific protein. The cellular ultrastructure of the melanocytes was observed by transmission electron microscopy. The cultured human melanocytes from face skin were multi-dendritic, and many mature melanosomes were observed. Therefore, using a specific culture medium, melanocytes from face skin can be successfully cultured and made available for further investigations.
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References
Abdel-Naser MB (2003) Mitogen requirements of normal epidermal human melanocytes in a serum and tumor promoter free medium. Eur J Dermatol 13:29–33
Akio M, Hajime I, Hideo O, Norio K (2004) Tyrosinase induction in normal human cultured melanocytes by endothelin-1. J Cardiovasc Pharmacol 44:S439–S442
Arita Y, Santiago-Schwarz F, Coppock DL (2000) Survival mechanisms induced by 12-O-tetradecanoylphorbol-13-acetate in normal human melanocytes include inhibition of apoptosis and increased Bcl-2 expression. Melanoma Res 10:412–420
Chen YF, Chang JS, Yang PY, Hung CM, Huang MH, Hu DN (2000) Transplant of cultured autologous pure melanocytes after laser-abrasion for the treatment of segmental vitiligo. J Dermatol 27:434–439
Davids LM, du Toit E, Kidson SH, Todd G (2009) A rare repigmentation pattern in a vitiligo patient: a clue to an epidermal stem-cell reservoir of melanocytes? Clin Exp Dermatol 34:246–248
Eisinger M, Marko O (1982) Selective proliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin. Proc Natl Acad Sci USA 79:2018–2022
Furuya R, Yoshida Y, Moro O, Tsunenaga M, Aoki H, Kishimoto J, Ifuku O, Hirobe T (2009) Immunohistochemical survey of the distribution of epidermal melanoblasts and melanocytes during the development of UVB-induced pigmented spots. J Dermatol Sci 55:99–107
Hirobe T (2001) Endothelins are involved in regulating the proliferation and differentiation of mouse epidermal melanocytes in serum-free primary culture. J Investig Dermatol Symp Proc 6:25–31
Kedlaya R, Kandala G, Liu TF, Maddodi N, Devi S, Setaluri V (2011) Interactions between GIPC-APPL and GIPC-TRP1 regulate melanosomal protein trafficking and melanogenesis in human melanocytes. Arch Biochem Biophys 508:227–233
Kumar R, Parsad D, Kanwar A, Kaul D (2012) Development of melanocye-keratinocyte co-culture model for controls and vitiligo to assess regulators of pigmentation and melanocytes. Indian J Dermatol Venereol Leprol 78:599–604
Munoz–Munoz JL, Acosta-Motos JR, Garcia-Molina F, Varon R, Garcia-Ruiz PA, Tudela J, Garcia-Canovas F, Rodriguez-Lopez JN (2010) Tyrosinase inactivation in its action on dopa. Biochim Biophys Acta 1804:1467–1475
Nielsen HI, Don P (1984) Culture of normal adult human melanocytes. Br J Dermatol 110:569–580
Oiso N, Tsuruta D, Imanishi H, Amatsu A, Kobayashi H, Kawara S, Kawada A (2008) Spotted hyperpigmentation: disfigured melanosomes in melanocytes and keratinocytes. J Eur Acad Dermatol Venereol 22:876–878
Petersson S, Shubbar E, Enerback L, Enerback C (2009) Expression patterns of S100 proteins in melanocytes and melanocytic lesions. Melanoma Res 19:215–225
Pittelkow MR, Shipley GD (1989) Serum-free culture of normal human melanocytes: growth kinetics and growth factor requirements. J Cell Physiol 140:565–576
Sulaimon SS, Kitchell BE (2003) The biology of melanocytes. Vet Dermatol 14:57–65
Swope VB, Medrano EE, Smalara D, Abdel-Malek ZA (1995) Long-term proliferation of human melanocytes is supported by the physiologic mitogens alpha-melanotropin, endothelin-1, and basic fibroblast growth factor. Exp Cell Res 217:453–459
Valtink M, Engelmann K (2007) Serum-free cultivation of adult normal human choroidal melanocytes. Graefes Arch Clin Exp Ophthalmol 245:1487–1494
Wang CC, Chen CK (2012) Effect of spot size and fluence on Q-switched alexandrite laser treatment for pigmentation in Asians: a randomized, double-blinded, split-face comparative trial. J Dermatol Treat 23:333–338
Zhu WY, Zhang RZ, Ma HJ, Wang DG (2004) Isolation and culture of amelanotic melanocytes from human hair follicles. Pigment Cell Res 17:668–673
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This work was supported by the CMA- L’OREAL CHINA SKIN GRANT 2010 (S2010070816).
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Tang, J., Li, Q., Cheng, B. et al. Primary culture of human face skin melanocytes for the study of hyperpigmentation. Cytotechnology 66, 891–898 (2014). https://doi.org/10.1007/s10616-013-9643-6
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DOI: https://doi.org/10.1007/s10616-013-9643-6