Summary
Two different dermal-epidermal recombinants were prepared in vitro and used to study the synthesis and formation of basement membrane. The first was obtained by culturing keratinocytes on the surface of a collagen lattice populated by fibroblasts. The second was prepared by coculture of both keratinocytes and fibroblasts in a collagen lattice. After 6 weeks of culture, the basal lamina was observed with electron microscopy only if keratinocytes were cultivated on top of the collagen lattice populated by fibroblasts. In the second model, however, type IV collagen, laminin, and pemphigoid bullosa antigen could be detected by immunofluorescence as well as synthesis of type IV collagen in the culture, but no basement membrane was observed by electron microscopy. These data demonstrate that, in vitro, basement membrane formation depends not only on the presence of the macromolecular components but also on the culture conditions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Asselineau D, Bernard B, Bailly C, Darmon M, Prunieras M (1986) Human epidermis reconstructed by culture: Is it “normal”? J Invest Dermatol 86:181–186
Bell E, Ivarsson B, Merrill C (1979) Production of a tissue like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential “in vitro.” Proc Natl Acad Sci USA 76:1274–1278
Bell E, Ehrlich P, Sher S, Merrill C, Sarber R, Hull B, Nakatsuji T, Church B, Buttle D (1981) Development and use of a living skin equivalent. Plast Reconstruct Surg 67:386–392
Bell E, Ehrlich P, Buttle D, Nakatsuji T (1981) Living tissue formed in vitro and accepted as skin equivalent tissue of full thickness. Science 211:1052–1054
Bell E, Sher S, Hull B, Merrill C, Rosen S, Chamson A, Asselineau D, Dubertret L, Coulomb B, Lapière C, Nusgens B, Neveux Y (1983) The reconstitution of living skin. J Invest Dermatol 81:2s-10s
Briggaman RA (1982) Epidermal-dermal interactions in adult skin. J Invest Dermatol 79:21s-29s
Briggaman RA, Dalldorf FG, Wheeler CE (1971) Formation and origin of basal lamina and anchoring fibrils in adult human skin. J Cell Biol 51:384–395
Chandrakajan G, Torchia DA, Piez KA (1976) Preparation of intact monomeric collagen from rat tail tendon and skin and the structure of the non helical ends in solution. J Biol Chem 251:6062–6067
Coulomb B, Dubertret L, Merrill C, Touraine R, Bell E (1984) The collagen lattice: a model for studying the physiology, biosynthetic function and pharmacology of the skin. Br J Dermatol 111:83–87
Coulomb B, Saiag P, Bell E, Breitburd F, Lebreton C, Heslan M, Dubretret L (1986) A new method for studying epidermalization in vitro. Br J Dermatol 114:91–101
David G, Nusgens B, Van der Schueren B, Van Cauwenberge D, Van der Berghe H, Lapière C (1987) Collagen metabolism and basement membrane formation in cultures of mouse mammary epithelial cells. Exp Cell Res 170:402–416
Eisinger MJ, Soo Lee Hefton JM, Darzynkiewicz Z, Chia JW, De Harven E (1979) Human epidermal cell cultures: growth and differentiation in the absence of dermal components or medium supplements. Proc Natl Acad Sci USA 76:5340–5344
Fuchs E, Green H (1981) Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell 25:617–625
Green H, Kehinde O, Thomas J (1979) Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. Proc Natl Acad Sci USA 76:5665–5668
Grinnell F, Takashima A, Lamke-Seymour C (1986) Morphological appearance of epidermal cells cultured on fibroblast reorganized collagen gels. Cell Tissue Res 246:13–21
Hirone T, Taniguchi S (1980) Basal lamina formation by epidermal cells in cell culture. In: Bernstein Seiji (ed) Biochemistry of normal and abnormal epidermal differentiation. Karger, Basel
Kitano E (1977) Studies on epidermo-dermal interactions with special reference to basal lamina formation. J Juzen Med Soc 86:286–303
Kleinman H, McGarvey M, Hassell J, Star V, Frances B, Laurie G, Martin GR (1986) Basement membrane complexes with biological activity. Biochemistry 25:312–318
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head for bacteriophage T4. Nature 227:680–685
Odland G, Ross R (1968) Human wound repair. I. Epidermal regeneration. J Cell Biol 39:135–151
Orkin R, Gehron RP, McGoodwin EB, Martin GR, Valentine T, Swarm R (1977) A murine tumor producing a matrix of basement membrane. J Exp Med 145:204–220
Périer C, Ronzière MC, Rattner A, Frey J (1980) Employment of gas liquid chromatography for the analysis of collagen amino acids in biopsy tissue. J Chromatogr 182:155–162
Pierce GB (1965) Basement membranes. VI. Synthesis by epithelial tumors of the mouse. Cancer 25:656–669
Régnier M, Desbas C, Bailly C, Darmon M (1988) Differentiation of normal and tumoral human keratinocytes cultured on dermis: reconstruction of either normal or tumoral architecture. In Vitro Cell Dev Biol 24:625–632
Saiag P, Coulomb B, Lebreton C, Bell E, Dubertret L (1985) Psoriatic fibroblasts induce hyperproliferation of normal keratinocytes in a skin equivalent model in vitro. Science 230:669–676
Sengel P (1976) Morphogenesis of skin. Cambridge University Press, Cambridge
Topol MB, Haimes BH, Dubertret L, Bell E (1986) Transfer of melanosomes in a skin equivalent model in vitro. J Invest Dermatol 87:642–647
Voller A, Bidwell DE, Bartlett A (1976) Enzyme immunoassays in diagnostic medicine. Bull WHO 53:55–65
Woodley D, Peterson H, Herzog S, Stricklin G, Burgeson R, Briggaman R, Cronce D, O'Keefe E (1988) Burn wounds resurfaced by cultured epidermal autografts show abnormal reconstitution of anchoring fibrils. Am Med Assoc 259:2566–2571
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chamson, A., Germain, N., Claudy, A. et al. Study of basement membrane formation in dermal-epidermal recombinants in vitro. Arch Dermatol Res 281, 267–272 (1989). https://doi.org/10.1007/BF00431061
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00431061