Influence of dermal equivalent maturation on the development of a cultured skin equivalent

Authors: Defrance-Bouvard, VéroniqueGermain, LucieRompré, PierreRoy, BrigitteAuger, François A.
Abstract: Histologie and Îmmunofluorescence methods were used to analyse the presence of fibronectin, chondroitin-4-sulphate and chondroitin-6-sulphate, type III and IV collagens, laminin, and keratins to assess the maturation level of cultured dermal and skin equivalents. ln a first phase, fibroblasts in monolayer culture were compared with dermal equivalents in which fibroblasts are embedded in a type 1 collagen gel. Different fluorescent patterns were observed depending on the culture system used. A sequential appearance of macromolecules was noticed in dermal equivalents. Fibronectin was first detected after 4 days of culture, whereas chondroitin-4-sulphate and chondroitin-6-sulphate and type III collagen were present after 7 days. ln cOntrast, aIl three macromolecules were detected at 24 h of culture in fibroblastic monolayer cultures. ln a second phase, the quality of our skin equivalents was evaluated according to the seeding time of epidermal cells upon dermal equivalents (1, 4, or 7 days). A satisfactory stratification was obtained when keratinocytes were seeded after 4 and 7 days of dermal equivalent culture. Laminin and fibronectin were detected at the dermo-epidermal junction, but type IV collagen was absent. Various keratins, as detected by the ABl, AB2, and AB3 antibodies, were present in the epidermallayer. Following keratinocyte confluence, a change in the organization pattern of type III collagen in the dermal fraction of the skin equivalent was also noticed. Our comparative results show that seeding of epidermal cells on a more mature dermal equivalent leads to improved differentiation status of the epidermallayer.
Document Type: Article de recherche
Issue Date: 1 January 1992
Open Access Date: Restricted access
Document version: VoR
This document was published in: Biochemistry and Cell Biology, Vol. 70 (1), 34–42 (1992)
National Research Council of Canada = Conseil national de recherches du Canada.
Alternative version: 10.1139/o92-005
Collection:Articles publiés dans des revues avec comité de lecture

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