Abstracts

Although canine skin models are already available as either monocellular or organotypic cultures, they only partly recapitulate normal skin organization and function.\udThe objective of this study was to establish a canine serum-free skin organ culture model and to test its pharmacological modulability.\udNormal skin was obtained from eight donor dogs referred for mastectomy. Biopsy samples were cultured in triplicate in Williams’E medium supplemented with penicillin/streptomycin, insulin, hydrocortisone and glutamine. Three experimental designs were performed: a) two-week viability of culture (N=2); b) Dexamethasone (DMS) inhibition of Epidermal Growth Factor (EGF) induced effects (N=3); c) Palmitoylethanolamide (PEA) down-modulation of compound 48/80 induced mast cell degranulation (N=3).\udGeneral morphological features of skin anatomical structures were well maintained up to day 14, scattered pyknotic nuclei were visible in the epidermis from day 7. Normal keratinocyte differentiation was confirmed by cytokeratin (K) 10, K14 and loricrin immunostaining. Epidermal thickness significantly decreased at day 14. Keratinocyte proliferation decrease was observed at day 7 and 14. Treatment with EGF induced both keratinocyte proliferation and thickening of epidermis that were counteracted by DMS. Compound 48/80 induced mast cell degranulation and the number of degranulated mast cells was reduced by PEA.\udCanine skin full thickness culture may offer unique opportunities to study skin pathophysiology and drugs mode of action in a biologically relevant 3D environment with the same three-dimensional cell–cell and cell–matrix contacts and communications present in the intact tissue