Abstract
The melanin pigment system in human skin is extraordinarly well developed and assures the photoprotection of the skin against harmful solar radiation. Specific cell-cell interactions between one melanocytes and keratinocytes play a fundamental role in the regulation of melanogenesis and melanin pigementation, the two key elements of this system, giving rise to the concept of a structural, functional collaborative ‘epidermal melanin unit,’ Early experiments strongly suggested that melanocyte growth and differentiation are regulated by paracrine factors from keratinocytes and other skin cells. In addition, co-culture studies with keratinocytes has shown that the extracellular matrix acts as a local environmental signal for dendrite formation and melanogenesis. Attempts to reconstruct pigmented human skin in vitro have made great progress over the last decade. The behavior of cells in these pigmented human skin equivalents closely resembles that in vivo, and the cells can still respond to appropriate extrinsic regulatory stimuli such as ultraviolet radiation. Keratinocytes and fibroblasts have been shown to be active partners in the regulation of melanocyte distribution, viability and other differentiation functions, presumably by direct contact and the effects of various soluble paracrine factors. By reproducing cell-cell and cell-matrix interactions, these culture systems provide a promising experimental model for investigating regulation of the skin pigmentary system and the role of photoprotection against harmful solar radiation.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Abdel-Malek, Z., Swope, V. B., Suzuki, I., Ackali, C., Harriger, M. D., Boyce, S. T., Urabe, K., andHearing, V. J. (1995): ‘Mitogenic and melanogenic stimulation of normal human melanocytes by melanotropic peptides,’Proc. Natl. Acad. Sci. USA,92, pp. 1789–1793
Archambault, M., Yaar, M., andGilchrest, B. A. (1995) ‘Keratinocytes and fibroblasts in a human skin equivalent model enhance melanocyte survival and melanin synthesis after ultraviolet irradiation,’J. Invest. Dermatol.,104, pp. 859–867
Asselineau, D., andPruniéras, M. (1984): ‘Reconstruction of “simplified” skin: control of fabrication,’Br. J. Dermatol.,111, pp. 219–222
Barker, D., Dixon, K., Medrano, E. E., Smalara, D., Im, S., Mitchell, D., Babcock, G., andAbdel-Malek, Z. A. (1995): ‘Comparison of the responses of human melanocytes with different melanin contents to UVB irradiation,’Cancer Res.,55, pp. 4041–4046
Bell, E., Ehrlich, H. P., Buttle, D. J., andNakatsuji, T. (1981): ‘Living tissue formedin vitro and accepted as skin-equivalent tissue of full thickness,’Science,211, pp. 1053–1054
Bell, E., Ivarsson, B., andMerrill, C. (1979): ‘Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potentialin vitro,’Proc. Natl. Acad. Sci. USA,76, pp. 1274–1278
Bertaux, B., Morliere, P., Moreno, G., Courtalon, A., Masse, J. M., andDubertret, L. (1988): ‘Growth of melanocytes in a skin equivalent modelin vitro,’Br. J. Dermatol.,119, pp. 503–512
Berthod, F., Hayek, D., Damour, O., andCollombel C. (1993): ‘Collagen synthesis by fibroblasts cultured within a collagen sponge,’Biomaterials,14, pp. 749–754
Bessou, S., Pain, C., andTaieb, A. (1997): ‘Use of human skin reconstructs in the study of pigment modifiers,’Arch. Dermat.,133, pp. 331–336
Bessou, S., Surleve Bazeille, J. E., Pain, C., Donatien, P., andTaieb, A. (1996): ‘Ex vivo study of skin phototypes,’J. Invest. Dermatol.,107, pp. 684–688
Boissy, R. E., Zhao, H., Oetting, W. S., Austin, L. M., Wildernberg S. C., Boissy, Y. L., Zhao, Y., Sturm, R. A., Hearing, V. J., King, R. A., andNordlund, J. J. (1996): ‘Mutation in and lack of expression of tyrosine-related protein-1 (TRP-1) in melanocytes from an individual with brown oculocutaneous albinism: a new subtype of albinism classified as “OCA3”,’Am. J. Hum. Genet.,58, pp. 1145–1156
Boyce, S. T., Medrano, E. E., Abdel, M. Z., Supp, A. P., Dodick, J. M., Nordlund, J. J., andWarden, G. D. (1993): ‘Pigmentation and inhibition of wound contraction by cultured skin substitutes with adult melanocytes after transplantation to athymic mice,’J. Invest. Dermatol.,100, pp. 360–365
Buffey, J. A., Messenger, A. G., Taylor, M., Ashcroft, A. T. T., Westgate, G. E., andMacNeil, S. (1994): ‘Extracellular matrix derived from hair and skin fibroblasts stimulates human skin melanocyte tyrosinase activity,’Br. J. Dermatol.,131, pp. 1–7
Chakraborty, A., Slominski, A., Ermak, G., Hwang, J., andPawelek, J. (1995): ‘Ultraviolet B and melanocyte-stimulating hormone (MSH) stimulate mRNA production for alphaMSH receptor and proopiomelanocortin-derived peptides in mouse melanoma cells and transformed kerationcytes,’J. Invest. Dermatol.,105, pp. 655–659
Chakraborty, A. K., Funasaka, Y., Slominski, A., Ermak, G., Hwang, J., Pawelek, J. M., andIchihashi, M. (1996): ‘Production and release of proopiomelanocortin (POMC) derived peptides by human melanocytes and keratinocytes in culture: regulation by ultraviolet B,’Biochim. Biophs. Acta.,1313, pp. 130–138
Chen, C. J., Lavker, R. M., Rodeck, U., Risse, B., andJensen, P. J. (1995): ‘Use of a serum-free epidermal culture medel to show deleterious effect of epidermal growth factor on morphogeneis and differentiation,’J. Invest. Dermatol.,104, pp. 107–112
Collombel, C., Damour, O., Gagnieu, C., Poinsignon, F., Echinard, C., andMarichy, J. (1987): ‘Peau artificielle et son procédé de fabrication,’ Brévet d'invention fransi no87-08752, 15 June 1987, Brévet d'invention europién no88-420194.8, 14 June 1988
Coulomb, B., Saiag, P., Bell, E., Breitburd, F., Lebreton, C., Heslan, M., andDubertret, L. (1986): ‘A new method for studying epidermalizationin vitro,’Br. J. Dermatol.,114, pp. 91–101
Daniels, F., Brophy, D., andLoditz, W. C. (1961): ‘Histological responses of human skin following ultraviolet irradiation,’J. Invest. Dermatol.,37, pp. 351–357
DeLuca, M., D'Anna, F., Bondanza, S., Franzi, A. T., andCancedda, R. (1988a): ‘Human epithelial cells induce human melanocyte growthin vitro but only skin keratinocytes regulate its proper differentiation in the absence of dermis,’J. Cell. Biol.,107, pp. 1919–1926
DeLuca, M., Franzi, A. T., D'Anna, F., Zicca, A., Albanese, E., Bondanza, S., andCancedda, R. (1988b): ‘Coculture, of human keratinocytes and melanocytes: differentiated melanocytes are physiologically ortganized in the basal layer of the cultured epithelium,’Eur. J. Cell. Biol.,46, pp. 176–180
DeLuca, M., Siegrist, W., Bondanza, S., Mathor, M., Cancedda, R., andEberle, A. N. (1993): ‘α-melanocyte stimulating hormon (α-MSH) stimulates normal melanocyte growth by binding to high affinity receptors,’J. Cell. Sci.,105, pp. 1079–1084
Derby, M. A. (1982): ‘Environmental factors affecting neural crest differentiation: melanocyte differentiation by crest cells exposed to cell-free (deoxycholate-exracted) dermal mesenchyme matrix,’Cell Tissue Res.,225, pp. 379–386
Eisinger, M., andMarko, 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, pp. 2018–2022
Eller, M. S., Ostrom, K., andGilchrest, B. A. (1996): ‘DNA damage enhaces melanogenesis,’Proc. Natl., Acad. Sci. USA,93, pp. 1087–1092
Eller, M. S., Yaar, M., andGilchrest, B. A. (1994): ‘DNA damage and melanogenesis,’Nature,372, pp. 413–414
Epstein, J. H. (1989): ‘Photocarcinogenesis, skin cancer and aging’InKligman, B. A. K., andKligman, A. M. (Eds.): ‘Aging and the skin’ (Raven Press, New York), pp. 307–349.
Fitzpatrick, T. B., andSzabo, G. (1959): ‘The melanocyte cytology and cytochemistry,’J. Invest. Dermatol.,32, pp. 197–209
Fitzpatrick, T. B., Szabo, S., andWick, M. M. (1983), ‘Biochemistry and physiology of melanin pigmentation,’inGoldsmith L. A. (Ed.) ‘Biochemistry and physiology of the skin. Vol. 2 (Oxford University Press), pp. 687–712
Freeman, A. E., Igel, H. J., Harrman, B.J., andKleinfield, K. L. (1976): ‘Growth and characterization of human skin epithelial cell culture,’In Vitro,12, pp. 352–362
Friedman, P. S., andGilchrest, B. A. (1987): ‘Ultraviolet radiation directly induces pigment production by cultured human melanocytes,’J. Cell. Physiol.,133, pp. 88–94
Funasaka, Y., Boulton, T., andCobb, M. (1992): ‘c-Kitkinase induces a cascade of protein tyrosine phosphorylation in normal human melanocytes in responses to mast cell growth factor and simulates mitogen-activated protein kinase but is down-regulated by melanosomes,’Mol. Biol. Cell,3, pp. 197–209
Fusenig, N. E. (1992): ‘Cell interaction and epithelial differentiation’inFreshney, R. I. (Ed.): ‘Culture of epithelial cells’ (Wiley-Liss, New York), pp. 25–57
Gardes, J., Lemke, H., Baisch, H., Wacker, H.-H., Schwab, U., andStein, H. (1984): ‘Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67,’J. Immunol.,133, pp. 1710–1715
Gilchrest, B. A., Albert, L. S., Karassik, R. L., andYaar, M. (1985): ‘Substrate influences human epidermal melanocyte attachment and spreadingin vitro,’In Vitro Cell. Dev. Biol.,21, pp. 114–120
Gilchrest, B. A., Zhai, S., Eller, M. S., Yarosh, D. B., andYaar, M. (1993): ‘Treatment of human melanocytes and S91 melanoma cell with the DNA repaire enzyme T4 endonuclease V enhances melanogenesis after ultraviolet irradiation,’J. Invest. Dermatol.,101, pp. 666–672
Haake, A. R., andScott, G. A. (1991): ‘Physiologic distribution and differentiation of melanocytes in human fetal and neonatal skin equivalents,’J. Invest. Dermatol.,96, pp. 71–77
Halaban, R., Langdon, R., Birchall, N., Cuono, C., Baird, A., Scott, G., Moellmann, G., andMcGuire, J. (1988): ‘Basic fibroblast growth factor from human keratinocytes is a natural mitogen for melanocytes,’J. Cell. Biol.,107, pp. 1611–1619
Halabane, R., Ghosh, S., andBaird, A. (1987): ‘bFGF is the putative natural growth factor for human melanocytes,’In Vitro Cell. Dev. Biol.,23, pp. 47–52
Halabane, R., Rubin, J. S., Funasaka, Y., Cobb, M., Boulton, T., Faletto, D., Rosen, E., Chan, A., Yoko, K., White, W., Cook, C., andMoellman, G. (1992): ‘Met and hepatocyte growth factor/scatter factor signal transduction in normal melanocytes and melanoma cells,’Oncogene,7, pp. 2195–2206
Hara, M., Yaar, M., andGilchrest, B. A. (1995): ‘Endothelin-1 of keratinocyte origin is a mediator of a melanocyte dendricity,’J. Invest. Dermatol.,105, pp. 744–748
Hedley, S., Gawkrodger, D. J., Weetman, A. P., andMacNeil, S. (1996): ‘Investigation of the influence of extracellular matrix proteins on normal human melanocyte tyrosinase activity,’Br. J. Dermatol.,135, pp. 888–897
Hedly, S., Wagner, M., Bielby, S., Smith-Thomas, L., Gawkrodger, D. J., andS. M. N. (1997): ‘The influence of extracellular matrix proteins on cutaneous and uveal melanocytes,’Pigment Cell. Res.,10, pp. 54–59
Herlyn, M., Mancianti, M., Jambrosic, J., Bolen, J. B., andKoprowski, H. (1988): ‘Regulatory factors that determine growth and phenotype of normal human melanocytes,’Exp. Cell. Res.,179, pp. 322–331
Imokawa, G., Yada, Y., andMiyagishi, M. (1992): ‘Endothelins secreted from keratinocytes are intrinsic mitogens for human melanocytes,’J. Biol. Chem.,267, pp. 24657–24680
Jimbow, K., Szabo, G., Pathak, M. A., andFitzpatrick, T. B. (1975) ‘Ultrastructural changes in human melanocytes after ultraviolet irradiation,inPathak, M. A. (Ed.): ‘Sunlight and man: normal and abnormal photobiologic responses’ (University of Tokyo Press), pp. 195–215
Knaggs, H. E., Holland, D. B., Morris, C., Wood, E. J., andCunliffe, W. J. (1994): ‘Quantification of cellular proliferation in acne using the monoclonal antibody Ki-67,’J. Invest. Dermatol.,102, pp. 89–92
Lavker, R. M., Gerberick, G. F., Veres, D., Irwin, C. J., andKaidbey, K. H. (1995): ‘Cumulative effects from repeated exposures to suberythermal doses of UVB and UVA in human skin’,J. Am. Acad. Dermatol.,32, pp. 53–62
McClenic, B. K., Mitra, R. S., Riser, B. L., Nickoloff, B. J., Dixit, V. M., andVarani, J. (1989): ‘Production and utilization of extracellular matrix components by human melanocytes,’Exp. Cell. Res.,180, pp. 314–25
Mishima, Y., andWidlen, S. (1967): ‘Enzymatically active and inactive melanocyte populations and ultraviolet irradiation: combined dopa-premelanin reactions and electron microscopy,’J. Invest. Dermatol.,49, 273–281
Morelli, G. J., Yohn, J. J., Lyons, M. B., Murphy, R. C., andNorris, D. A. (1989): ‘Leukotrienes C4 and D4 as potent mitogens for cultured human neonatal melanocytes,’J. Invest. Dermatol.,93, pp. 719–722
Nakazawa, K., Nakazawa, H., Collombel, C., andDamour, O. (1995): ‘Keratinocyte extracellular matrix-mediated regulation of normal human melanocyte functions,’Pigment Cell Res.,8, 10–18
Nakazawa, K., Nakazawa, H., Sauc, F., Lepavec, A., Collombel, C., andDamour, O. (1997): ‘Pigmented human skin equivalent—mew method of reconstitution by grafting an epithelial sheet onto a non-contractile dermal equivalent,’Pigment Cell Res.
Naughton, G. K., Jacob, L., andNaughton, B. A. (1989): ‘A physiological skin model for in vitro toxicity studies. InA. M. Goldberg (Ed.) ‘In vitro toxicology new directions. Vol. 7’ (Mary Ann Liebert Inc., New York), pp. 183–189
Nordlund, J. J., Collins, C. E., andRheins, L. A. (1986): ‘Prostaglandin E2 and D2 but not MSH stimulate the proliferation of pigment cells in the pinnal epidermis of the DBA/2 mouse,’J. Invest. Dermatol.,86, pp. 433–437
Pathak, M. A., Sinesi, S. J., andSzabo, G. (1965): ‘The effect of a single dose of ultraviolet radiation on epidermal melanocytes,’J. Invest. Dermatol.,45, pp. 520–528
Pittelkow, M. R., andShipley, G. D. (1989): ‘Serum-free culture of normal human melanocytes: growth kinetics and growth factor requirements,’J. Cell. Physiol.,140, pp. 565–576
Pruniéras, M., Régnier, M., andWoodley, D. (1983): ‘Methods for cultivation of keratinocytes with an air-liquid interface,’J. Invest. Dermatol.,81, pp. 28s-33s
Quevedo, W. C., Szabo, G., Virks, J., andSinesi, S. J. (1965): ‘Melanocypte populations in UV-irradiated human skin,’J. Invest. Dermatol.,45, pp. 295–298
Ranson, M., Posen, S., andMason, R. S. (1988): ‘Extracellular matrix modulates the function of human melanocytes but not melanoma cells,’J. Cell. Physiol.,136, pp. 281–288
Rosdahl, I. K., andSzabo, G. (1978): ‘Mitotic activity of epidermal melanocytes in UV-irradiated mouse skin,’J. Invest. Dermatol.,70, pp. 143–148
Rosen, C. F., Seki, Y., Farinelli, W., Stern, R. S., Fitzpatrick, T. B., Pathak, M. A., andGange, R. W. (1987): ‘A comparison of the melanocyte response to narrow band UVA and UVB exposurein vivo,’J. Invest. Dermatol.,88, pp. 774–779
Schauer, E., Trautinger, F., Kock, A., Schwartz, A., Bhardwaj, R., Simon, M., Ansel, J. C., Schwarz, T., andLuger, T. A. (1994): ‘Proopiomelanocortin-derived peptides are synthesized and released by human keratinocytes,’J. Clin. Invest.,93, 2258–2262
Scott, G., Liang, H., andLuthra, D. (1996): ‘Stein cell factor regulates the melanocyte cytoskeleton,’Pigment Cell Res.,9, pp. 134–141
Scott, G., Ryan, D. H., andMcCarthy, J. B. (1992): ‘Molecular mechanisms of human melanocyte attachment to fibronectin,’J. Invest. Dermatol.,99, pp. 787–794
Scott, G. A., andHaake, A. R. (1991): ‘Keratinocytes regulate melanocyte number in human fetal and neontal skin equivalents,’J. Invest. Dermatol.,97, pp. 776–781
Scott, G. C. L., andBusacco, A. (1997): ‘Fibronectin suppresses apoptosis in normal human melanocytes through an integrin-dependent mechanism,’J. Invest. Dermatol.,108, pp. 147–153 (1997)
Slominski, A., Moellmann, G., Kuklinska, E., Bomirski, A., andPawelek, J. (1988): ‘Positive regulation of melanin pigmentation by two key substrates of the melanogenic pathway, L-tyrosine and L-dopa,’J. Cell. Sci.,89, pp. 287–296
Snell, R. S. (1963): ‘The effect of ultraviolet irradiation on melanogenesis,’J. Invest. Dermatol.,40, pp. 127–132
Soini, Y, Kamel, D., Paakko, P., Lehto, V. P., Oikarinen, A., andVahakangas, K. (1994): ‘Aberrant accumulation of p53 associates with Ki67 and mitotic count in benign skin legions,’Br. J. Dermatol.,131, pp. 514–520
Stierner, U., Rosdahl, I., Augustsson, A., andKagedal, B. (1989): ‘UVB irradiation induces melanocyte increase in both exposed and shielded human skin,’J. Invest. Dermatol.,92, pp. 561–564
Swope, V. B., Abdel-Malek, Z., Kassem, L. M., andNordlund, J. J. (1991): ‘Interleukins 1 alpha and 6 and tumor necrosis factoralpha are paracrine inhibitors of human melantocyte proliferation and melanogenesis,’J. Invest. Dermatol.,96, pp. 180–185
Tobin, D., Quinn, A. G., Ito, S., andThody, A. J. (1994): ‘The presence of tyrosinase and related proteins in human epidermis and their relationship to melanin type,’Pigment Cell Res.,7, pp. 204–209
Todd, C., Hewitt, S. D., Hempenaar, J., Noz, K., Thody, A. J., andPonec, M. (1993): ‘Co-culture of human melanocytes and keratinocytes in a skin equivalent model: effect of ultraviolet radiation,’Arch. Dermatol. Res.,285, pp. 455–459
Tomita, Y, Iwamoto, M., Masuda, T., andTagami, H. (1987): ‘Stimulatory effect of prostaglandin E2 on the configuration of normal human melanocytesin vitro’J. Invest. Dermatol.,89, pp. 299–301
Tomita, Y., Torinuki, W., andTagmi, H. (1988): ‘Stimulation of human melanocytes by Vitamin D3 possibly mediates skin pigmentation after sun exposure,’J. Invest. Dermatol.,90, pp. 882–884
Topol, B. M., Haimes, H. B., Dubertret, L., andBell, E. (1986): ‘Transfer of melansomes in a skin equivalent modelin vitro,’J. Invest. Dermatol.,87, pp. 642–647
Tsunenaga, M., Kohno, Y., Horii, I., Yasumoto, S., Huh, N. H., Tackikawa, T., Yoshiki, S., andKuroki, T. (1994): ‘Growth and differentiation properties of normal and transformed human keratinocytes in organotypic culture,’Jpn. J. Cancer Res.,85, pp. 238–244
Valyi, N. I., andHerlyn, M. (1991) ‘Regulation of growth and phenotype of normal human melanocytes in culture,’Cancer Treat. Res.,54, pp. 85–101
Vijayasaradhi, S., Doskoch, P. M., Wolchok, J., andHoughton, A. N. (1995): ‘Melanocyte differentiation marker gp75, the brown locus protein, can be regulated independently of tyrosinase and pigmentation,’J. Invest. Dermatol.,105, pp. 113–119
Wilkins, L., Gilchrest, B. A., Szabo, G., Weinstein, R., andMaciag, T. (1985): ‘The stimulation of normal human melanocyte proliferationin vitro by melanocyte growth factor from bovine brain,’J. Cell Physiol.,122, pp. 350–61
Yaar, M., andGilchrest, B. A. (1991): ‘Human melanocyte growth and differentiation: a decade of new data,’J. Invest. Dermatol.,97, pp. 611–617
Yaar, M., Grossman, K., Eller, M., andGilchrest, B. A. (1991): ‘Evidence for nerve growth factior-mediated paracrine effects in human epidermis,’J. Cell Biol.,115, pp. 821–828
Yada, Y., Higuchi, K., andImokawa, G. (1991): ‘Effects of endothelins on signal transduction and proliferation in human melanocytes,’J. Biol. Chem.,266, pp. 18352–18357
Yannas, I. V., andBurke, J. F. (1980): ‘Design of the artificial skin I. Basic design principals,’J. Biomed. Mat. Res.,14, pp. 65–81
Yohn, J. J., Morelli, J. G., Walchak, S. J., Rundell, K. B., Norris, D. A., andZamora, M. R. (1993): ‘Cultured human keratinocytes synthesize and secrete endothelin-1,’J. Invest. Dermatol.,100, pp. 23–26
Yuspa, S. H., andDlugosz, A. A. (1991): ‘Cutaneous carcinogenesis: natural and experimental’inGoldsmith, L. A. (Ed.): ‘Physiology biochemistry and molecular biology of the skin’ (Oxford University Press), pp. 1365–1402
Zhao, H., Eling, D. J., Medrano, E. E., andBoissy, R. E. (1996): ‘Retroviral infection with human tyrosinase-related protein-1 (TRP-1) cDNA upregulates tyrosinase activity and melanin synthesis in a TRP-1-deficient melanoma cell line,’J. Invest. Dermatol.,106, pp. 744–752
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nakazawa, K., Kalassy, M., Sahuc, F. et al. Pigmented human skin equivalent—as a model of the mechanisms of control of cell-cell and cell-matrix interactions. Med. Biol. Eng. Comput. 36, 813–820 (1998). https://doi.org/10.1007/BF02518888
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02518888