BROADENING THE APPLICATION OF CULTURED HUMAN KERATINOCYTE GRAFTING

Abstract

This thesis describes a body of work aimed at setting up and testing an in vivo experimental system, in which differentiated human epidermis can be used in laboratory experiments in experiments that could last several weeks. A series of experiments was carried out to establish a system for generating long-lived, fully-differentiated human skin patches on immunodeficient mouse hosts. Finally, the optimized systems were combined to generate mature grafts using keratinocytes carrying genetic disease mutants, and the grafts were tested for their ability to reproduce the disease phenotype. It is concluded that this model of human-to-mouse xenografts of culture generated human skin equivalent constructs, can indeed be applied to mutant keratin EBS-mimetic cells to imitate the EBS patient’s disorder, as determined by internal structural damage in keratinocytes. However, assays to show EBS-like tissue disruption will require a more mature, older graft. These findings are important for the development of tissue-relevant preclinical testing, because they lower the risk of expensive drug trial failures.