A versatile murine 3D organotypic model to evaluate aspects of wound healing and epidermal organization
Kandyba E, Hodgins M, Martin P.
Department of Biological and Biomedical Sciences, School of Life Sciences, Glasgow Caledonian University
Methods in Molecular Biology; Epidermal Cells (Book)
Three-dimensional (3D) organotypic models are increasingly being used to study aspects of epidermal organisation and cutaneous wound-healing events. These are largely dependent on laborious histological analysis and immunohistochemical approaches. Here we outline a method for establishment of a versatile in vitro 3D organotypic skin equivalent that reflects murine epidermal organisation in vivo. The system is optically transparent and ideally suited to real-time analysis and integrated in situ imaging techniques. Moreover, the model permits the visualisation of epidermal regeneration following injury in real time, thereby facilitating avenues to explore distinctive modes of wound re-epithelialisation. The versatility of the model could help unravel molecular mechanisms underlying epidermal morphogenesis, assess novel therapeutic strategies and reduce animal experimentation in a non-invasive manner.
Protocol for murine three-dimensional (3D) organotypic skin equivalent, confocal immunofluorescence and wound assay. CnT-07 was called Progenitor cell targeted (PCT) keratinocyte medium, and referred to as keratinocyte growth medium (KGM).