Summary
Merkel cells constitute a unique population that remains difficult to characterize in human skin because of their scarcity. Our aim was to develop tools for the study of Merkel cells in vitro. As a first step, we evaluated the possibility of harvesting human Merkel cells with the two-step extraction method that is widely used to extract and culture keratinocytes. Merkel cells were identified in the epithelial portion of hairy or glabrous skin biopsies by keratin (K)18 and K20 labeling. The totality of cutaneous epithelial cells were isolated from either hairy or glabrous skin biopsies following enzymatic dissociation of both the epidermis and the hair follicles. Flow cytometry was performed to quantify the small Merkel cell population. The analysis revealed that K18-labeled cells represented between 4.0 and 7.6% of freshly dissociated basal epidermal cells. No significant differences were seen between samples derived from glabrous palmar and hairy anatomic sites from children and adults, respectively. We also reported on the presence of Merkel cells in primary and first subcultures of human epidermal cells. The next step will be to enrich the isolated human Merkel cells and improve their culture conditions. An amplification of the number of Merkel cells will allow further studies to unravel long-standing questions regarding their origin, proliferative capacity, and functions in cutaneous biology.
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References
Auger FA, Remy-Zolghadri M, Grenier G, Germain L (2000) The self-assembly approach for organ reconstruction by tissue engineering. Biomed J Regenerative Med 1: 75–86
Compton CC, Regauer S, Seiler GR, Landry DB (1990) Human Merkel cell regeneration in skin derived from cultured keratinocyte grafts. Lab Invest 62: 233–2241
Fradette J, Godbout M-J, Michel M, Germain L (1995) Localization of Merkel cells at hair- less and hairy human skin sites using keratin 18. Biochem Cell Biol 73: 635–639
Fradette J, Larouche D, Fugère C, Guignard R, Beauparlant A, Couture V, CaouetteLaberge L, Roy A, Germain L (2003) Normal human Merkel cells are present in epidermal cell populations isolated and cultured from glabrous and hairy skin sites. J Invest Dermatol 120 (2): 313–317
Germain L, Rouabhia M, Guignard R, Carrier L, Bouvard V, Auger FA (1993) Improve- ment of human keratinocyte isolation and culture using thermolysin. Burns 2: 99–104
Germain L, Michel M, Fradette J, Xu W, Godbout M-J, Li H (1997) Skin stem cell identification and culture: a potential tool for rapid epidermal sheet production and grafting. In: Rouabhia M (ed) Skin substitute production by tissue engineering: clinical and fundamental applications. Landes Bioscience, Austin, TX, pp 177–210
Green H, Kehinde O, Thomas J (1979) Growth of cultured human epidermal cells into a multiple epithelia suitable for grafting. Proc Natl Acad Sci USA 76: 5665–5668
Kaufmann R, Frosch D, Westphal C, Weber L, Klein C (1989) Integrin VLA-3: ultrastructural localization at cell-cell contact sites of human cell cultures. J Cell Biol 109: 18071815
Lacour JP, Dubois D, Pisani A, Ortonne JP (1991) Anatomical mapping of Merkel cells in normal human adult epidermis. Br J Dermatol 125: 535–542
McKenna Boot P, Rowden G, Walsh N (1992) The distribution of Merkel cells in human fetal and adult skin. Am J Dermatopathol 14: 391–396
Michel M, Török N, Godbout M-J, Lussier M, Gaudreau P, Royal A, Germain L (1996) Keratin 19 as a biochemical marker of skin stem cells in vivo and in vitro: keratin 19 expressing cells are differentially localized in function of anatomic sites, and their number varies with donor age and culture stage. J Cell Sci 109: 1017–1028
Moll I (1994) Merkel cell distribution in human hair follicles of the fetal and adult scalp. Cell Tissue Res 277: 131–138
Moll I, Kuhn C, Moll R (1995) Cytokeratin 20 is a general marker of cutaneous Merkel cells while certain neuronal proteins are absent. J Invest Dermatol 104: 910–915
Moll R, Moll I, Franke WW (1984) Identification of Merkel cells in human skin by specific cytokeratin antibodies: changes of cell density and distribution in fetal and adult plantar epidermis. Differentiation 28: 136–154
Narisawa Y, Hashimoto K, Nakamura Y, Kohda H (1993) A high concentration of Merkel cells in the bulge prior to the attachment of the arrector pili muscle and the formation of the perifollicular nerve plexus in human fetal skin. Arch Dermatol Res 285: 261–268
Narisawa Y, Hashimoto K, Kohda H (1994) Merkel cells of the terminal hair follicle of the adult human scalp. J Invest Dermatol 102: 506–510
Rheinwald JG, Green H (1975) Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6: 331–343
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Fradette, J. et al. (2003). Dissociation, Quantification and Culture of Normal Human Merkel Cells Among Epidermal Cell Populations Derived from Glabrous and Hairy Skin Sites. In: Baumann, K.I., Halata, Z., Moll, I. (eds) The Merkel Cell. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-10358-6_17
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DOI: https://doi.org/10.1007/978-3-662-10358-6_17
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-05574-4
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