title: Standardized, defined serum-free culture of a human skin equivalent on fibroblast-populated collagen scaffold.
abstract: The defined, serum-free media used in the cultivation of skin equivalents are liable to inter-laboratory variations, require the preparation of multiple additives, and are potentially difficult to replicate. In this study, we assessed the use of standardized, serum-free and bovine pituitary extract-free keratinocyte culture media in the development of a human skin equivalent. After culture at the air-liquid interface for 3 weeks on a fibroblast-populated collagen lattice, an orthokeratinized, pluristratified epithelium was produced which expressed cytokeratins, cornified cell envelope precursors (involucrin, transglutaminase 1, filaggrin) and desmosomal components (desmoglein and desmocollin 1 and 3, plakophilin 1) in a differentiation-specific manner. There was also evidence of basement membrane reconstitution with collagen IV/VII, laminin 5, α6 and β4 integrin subunit expression at the epithelial-matrix junction. Overall, our findings demonstrate that readily available, defined organotypic culture media can be used to generate a reproducible skin equivalent with hallmarks of epidermal differentiation.
authors: Department of Dermatology and Atopy Research Centre, Juntendo University Graduate School of Medicine, Tokyo, Japan. wng@juntendo.ac.jp
authors: William
authors: W
authors: Ng
authors: Shigaku
authors: S
authors: Ikeda
mesh_terms: D001485:Basement Membrane / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000592:standards; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism*; D016895:Culture Media, Serum-Free / Q000592:standards; D003896:Desmosomes / Q000378:metabolism; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D000091344:Filaggrin Proteins; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism; D017768:Skin, Artificial; D013997:Time Factors; D054457:Tissue Scaffolds
chemical_list: D015415:Biomarkers; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007633:Keratins; D003094:Collagen