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Establishment and pharmacological modulation of a canine organ culture model

机译：犬器官培养模型的建立与药理调控

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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.
机译：尽管犬皮肤模型已经可以通过单细胞或器官型培养获得，但它们只能部分概括正常的皮肤组织和功能。\ ud本研究的目的是建立犬无血清皮肤器官培养模型并测试其药理学可调节性。\ ud正常皮肤是从八只进行乳房切除术的供体犬获得的。将活检样品在补充有青霉素/链霉素，胰岛素，氢化可的松和谷氨酰胺的威廉姆斯E培养基中一式三份培养。进行了三个实验设计：a）培养两周的生存力（N = 2）； b）地塞米松（DMS）抑制表皮生长因子（EGF）诱导的作用（N = 3）； c）化合物48/80诱导的肥大细胞脱粒（N = 3）的棕榈酰乙醇酰胺（PEA）下调。直到第14天，皮肤解剖结构的一般形态特征都得到了很好的维持，从表皮开始可见散在的表皮结核。 7.细胞角蛋白（K）10，K14和loricrin免疫染色证实了正常的角质形成细胞分化。在第14天，表皮厚度显着降低。在第7天和第14天，观察到角质形成细胞增殖减少。用EGF处理诱导角质形成细胞增殖和表皮增厚，这被DMS所抵消。化合物48/80诱导的肥大细胞脱粒并通过PEA减少了肥大细胞的脱颗粒。\ ud犬的皮肤全层培养可能会提供独特的机会来研究在生物学相关的3D环境中使用相同的3D环境进行皮肤病理生理学和药物作用方式完整组织中存在的维细胞间和细胞基质接触与通信。

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