Archives of Dermatological Research

, Volume 281, Issue 4, pp 267–272 | Cite as

Study of basement membrane formation in dermal-epidermal recombinants in vitro

  • A. Chamson
  • N. Germain
  • A. Claudy
  • C. Perier
  • J. Frey
Original Contributions


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.

Key words

Basement membrane Dermal-epidermal recombinants Keratinocyte culture Collagen Dermal equivalent 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Asselineau D, Bernard B, Bailly C, Darmon M, Prunieras M (1986) Human epidermis reconstructed by culture: Is it “normal”? J Invest Dermatol 86:181–186Google Scholar
  2. 2.
    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 76:1274–1278Google Scholar
  3. 3.
    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 67:386–392Google Scholar
  4. 4.
    Bell E, Ehrlich P, Buttle D, Nakatsuji T (1981) Living tissue formed in vitro and accepted as skin equivalent tissue of full thickness. Science 211:1052–1054Google Scholar
  5. 5.
    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-10sGoogle Scholar
  6. 6.
    Briggaman RA (1982) Epidermal-dermal interactions in adult skin. J Invest Dermatol 79:21s-29sGoogle Scholar
  7. 7.
    Briggaman RA, Dalldorf FG, Wheeler CE (1971) Formation and origin of basal lamina and anchoring fibrils in adult human skin. J Cell Biol 51:384–395Google Scholar
  8. 8.
    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 251:6062–6067Google Scholar
  9. 9.
    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 111:83–87Google Scholar
  10. 10.
    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 114:91–101Google Scholar
  11. 11.
    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 170:402–416Google Scholar
  12. 12.
    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 76:5340–5344Google Scholar
  13. 13.
    Fuchs E, Green H (1981) Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell 25:617–625Google Scholar
  14. 14.
    Green H, Kehinde O, Thomas J (1979) Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. Proc Natl Acad Sci USA 76:5665–5668Google Scholar
  15. 15.
    Grinnell F, Takashima A, Lamke-Seymour C (1986) Morphological appearance of epidermal cells cultured on fibroblast reorganized collagen gels. Cell Tissue Res 246:13–21Google Scholar
  16. 16.
    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, BaselGoogle Scholar
  17. 17.
    Kitano E (1977) Studies on epidermo-dermal interactions with special reference to basal lamina formation. J Juzen Med Soc 86:286–303Google Scholar
  18. 18.
    Kleinman H, McGarvey M, Hassell J, Star V, Frances B, Laurie G, Martin GR (1986) Basement membrane complexes with biological activity. Biochemistry 25:312–318Google Scholar
  19. 19.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head for bacteriophage T4. Nature 227:680–685Google Scholar
  20. 20.
    Odland G, Ross R (1968) Human wound repair. I. Epidermal regeneration. J Cell Biol 39:135–151Google Scholar
  21. 21.
    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 145:204–220Google Scholar
  22. 22.
    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 182:155–162Google Scholar
  23. 23.
    Pierce GB (1965) Basement membranes. VI. Synthesis by epithelial tumors of the mouse. Cancer 25:656–669Google Scholar
  24. 24.
    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 24:625–632Google Scholar
  25. 25.
    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 230:669–676Google Scholar
  26. 26.
    Sengel P (1976) Morphogenesis of skin. Cambridge University Press, CambridgeGoogle Scholar
  27. 27.
    Topol MB, Haimes BH, Dubertret L, Bell E (1986) Transfer of melanosomes in a skin equivalent model in vitro. J Invest Dermatol 87:642–647Google Scholar
  28. 28.
    Voller A, Bidwell DE, Bartlett A (1976) Enzyme immunoassays in diagnostic medicine. Bull WHO 53:55–65Google Scholar
  29. 29.
    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 259:2566–2571Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • A. Chamson
    • 1
  • N. Germain
    • 1
  • A. Claudy
    • 2
  • C. Perier
    • 1
  • J. Frey
    • 1
  1. 1.Biochemistry DepartmentMedical SchoolSaint-Etienne Cedex 2France
  2. 2.Dermatology DepartmentMedical SchoolSaint-Etienne Cedex 2France

Personalised recommendations