Generation of a Full-Thickness Human Skin Equivalent on an Immunodeficient Mouse
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Human skin equivalents composed of epidermal cells and fibroblasts are important for modeling human epidermal development, testing new therapeutics, and designing novel treatment strategies for human skin diseases. Here, we describe a procedure for the generation of an in vivo full-thickness human skin equivalent on an immunodeficient mouse using a grafting chamber system. The protocol involves mixing human epidermal cells and fibroblasts in a silicone grafting chamber that is surgically inserted onto the muscle fascia of a recipient immunodeficient mouse. Following the removal of the silicone chamber, the graft area is exposed to air to induce stratification of developing epidermis, resulting in the reconstitution of full-thickness human skin tissue on a live mouse. This grafting system provides a straightforward approach to study human skin diseases in an animal model and has been previously used to determine the ability of both mouse and human primary epidermal cells and cells derived from pluripotent stem cells to regenerate functional skin in vivo.
KeywordsXenograft Human skin graft Mouse model Grafting chamber Keratinocytes Epidermal cells Fibroblasts
Hematoxylin and eosin
Nonobese diabetic/severe combined immunodeficiency
We are grateful for the funding support from the National Institutes of Health (T32 AR007411-33) and the University of Colorado Skin Diseases Research Core Center (P30 AR057212). We also thank Epidermolysis Bullosa (EB) Research Partnership, the EB Medical Research Foundation, the Cure EB Charity, Dystrophic Epidermolysis Bullosa Research Association (DEBRA) International, and the Gates Frontiers Fund.
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