A new method for studying epidermalization in vitro |
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Authors: | B. COULOMB P. SAIAG E. BELL F. BREITBURD C. LEBRETON M. HESLAN L. DUBERTRET |
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Affiliation: | Laboratoire de Dermatologie, Service de Dermatologie, Hopital Henri Mondor, Crétcil, France;Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, U.S.A.;Unitédes Papillomavirus, EU 190, Institut Pasteur, Paris, France |
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Abstract: | A new method for studying epidermalization in vitro is described. It consists of inserting a punch biopsy that serves as a source of epidermis into dermal equivalent freshly made up, with fibroblasts mixed in a collagen matrix. Fibroblasts cling to collagen fibrils and contract the matrix, leading in 3 days to a resistant dermal equivalent holding the punch biopsy firmly in place. At day 5, a culture medium favouring epidermal growth was used and a fringe of a new epidermis appeared around the punch, the area of which grew linearly with time. This new epidermis showed a pattern of differentiation similar to epidermis in vivo, with cuboidal basal cells, keratohyalin granules, membrane coating granules and the expression of the 65-67 kd keratin subset. The method seems to combine the advantages of the explant technique and of classical keratinocyte cultures, providing the researcher with a large quantity of differentiated epidermis, the pharmacologist with simple and quantitative system in which to study modifications of growth and differentiation of epidermis, and the plastic surgeon with a possible material for skin grafting. |
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