期刊
ACTA DERMATO-VENEREOLOGICA
卷 91, 期 4, 页码 387-391出版社
ACTA DERMATO-VENEREOLOGICA
DOI: 10.2340/00015555-1092
关键词
serum-free media; skin substitute; organotypic co-culture
类别
资金
- Ministry of Education, Culture, Sport, Science and Technology (MEXT) of the Japan Government [S0991013]
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, alpha 6 and beta 4 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.
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