Abstract
Melanocytes which represent around 5% of epidermal cells are located in the basal layer. To culture melanocytes we used trypsin digestion instead of dispase to obtain a cell suspension containing only basal keratinocytes and melanocytes. Melanocytes are cells which need a great attention. Indeed they dedifferentiate easily in culture as soon as they are in pure culture. Factors secreted by contaminating keratinocytes allow melanocytes to stay dendritic but by regulating their number avoid their growth. In order to age, phototype and other individual dependent factors regulate the behavior of melanocytes in vitro. Thus, microscopic examination of melanocytes has to be performed each day to adapt conditions of culture to each primary cell culture. This is the secret to have a nice melanocyte culture.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bonaventure J, Domingues MJ, Larue L (2013) Cellular and molecular mechanisms controlling the migration of melanocytes and melanoma cells. Pigment Cell Melanoma Res 26(3):316–325
Haass NK, Herlyn M (2005) Normal human melanocyte homeostasis as a paradigm for understanding melanoma. J Investig Dermatol Symp Proc 10(2):153–163
Jimbow K, Quevedo WC, Fitzpatrick TB, Szabo G (1976) Some aspects of melanin biology: 1950–1975. J Invest Dermatol 67(1):72–89
Nishimura EK (2011) Melanocyte stem cells: a melanocyte reservoir in hair follicles for hair and skin pigmentation. Pigment Cell Melanoma Res 24(3):401–410
Tadokoro T, Yamaguchi Y, Batzer J, Coelho SG, Zmudzka BZ, Miller SA et al (2005) Mechanisms of skin tanning in different racial/ethnic groups in response to ultraviolet radiation. J Invest Dermatol 124(6):1326–1332
Démarchez M. Melanocyte and pigmentation. 2011. https://biologiedelapeau.fr/spip.php?article7
Szabo G (1967) The regional anatomy of the human integument with special reference to the distribution of hair follicles, sweat glands and melanocytes. Philos Trans R Soc Lond Ser B Biol Sci 252(779):447–485
Yamaguchi Y, Itami S, Watabe H, Yasumoto K-I, Abdel-Malek ZA, Kubo T et al (2004) Mesenchymal-epithelial interactions in the skin: increased expression of dickkopf1 by palmoplantar fibroblasts inhibits melanocyte growth and differentiation. J Cell Biol 165(2):275–285
Fitzpatrick TB, Breathnach AS (1963) The epidermal melanin unit system. Dermatol Wochenschr 147:481–489
Hennessy A, Oh C, Diffey B, Wakamatsu K, Ito S, Rees J (2005) Eumelanin and pheomelanin concentrations in human epidermis before and after UVB irradiation. Pigment Cell Res 18(3):220–223
Hunt G, Kyne S, Ito S, Wakamatsu K, Todd C, Thody A (1995) Eumelanin and phaeomelanin contents of human epidermis and cultured melanocytes. Pigment Cell Res 8(4):202–208
Tobin D, Quinn AG, Ito S, Thody AJ (1994) The presence of tyrosinase and related proteins in human epidermis and their relationship to melanin type. Pigment Cell Res 7(4):204–209
Quevedo WC, Fitzpatrick TB, Pathak MA, Jimbow K (1975) Role of light in human skin color variation. Am J Phys Anthropol 43(3):393–408
Cook AL, Donatien PD, Smith AG, Murphy M, Jones MK, Herlyn M et al (2003) Human melanoblasts in culture: expression of BRN2 and synergistic regulation by fibroblast growth factor-2, stem cell factor, and endothelin-3. J Invest Dermatol 121(5):1150–1159
Hirobe T (1992) Melanocyte stimulating hormone induces the differentiation of mouse epidermal melanocytes in serum-free culture. J Cell Physiol 152(2):337–345
Zhao Z, Jin C, Ding K, Ge X, Dai L (2012) Dedifferentiation of human epidermal melanocytes into melanoblasts in vitro. Exp Dermatol 21(7):504–508
Kormos B, Belső N, Bebes A, Szabad G, Bacsa S, Széll M et al (2011) In vitro dedifferentiation of melanocytes from adult epidermis. PLoS One 6(2):e17197
Kustikova OS, Wahlers A, Kühlcke K, Stähle B, Zander AR, Baum C et al (2003) Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population. Blood 102(12):3934–3937
Hachiya A, Kobayashi A, Ohuchi A, Takema Y, Imokawa G (2001) The paracrine role of stem cell factor/c-kit signaling in the activation of human melanocytes in ultraviolet-B-induced pigmentation. J Invest Dermatol 116(4):578–586
Luo D, Chen H, Searles G, Jimbow K (1995) Coordinated mRNA expression of c-Kit with tyrosinase and TRP-1 in melanin pigmentation of normal and malignant human melanocytes and transient activation of tyrosinase by Kit/SCF-R. Melanoma Res 5(5):303–309
Hirobe T, Osawa M, Nishikawa S-I (2004) Hepatocyte growth factor controls the proliferation of cultured epidermal melanoblasts and melanocytes from newborn mice. Pigment Cell Res 17(1):51–61
Sviderskaya EV, Wakeling WF, Bennett DC (1995) A cloned, immortal line of murine melanoblasts inducible to differentiate to melanocytes. Development 121(5):1547–1557
Holbrook KA, Underwood RA, Vogel AM, Gown AM, Kimball H (1989) The appearance, density and distribution of melanocytes in human embryonic and fetal skin revealed by the anti-melanoma monoclonal antibody, HMB-45. Anat Embryol (Berl) 180(5):443–455
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Cario, M., Taieb, A. (2019). Isolation and Culture of Epidermal Melanocytes. In: Böttcher-Haberzeth, S., Biedermann, T. (eds) Skin Tissue Engineering. Methods in Molecular Biology, vol 1993. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9473-1_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9473-1_3
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9472-4
Online ISBN: 978-1-4939-9473-1
eBook Packages: Springer Protocols