Intractable skin ulcers resulting from diabetes, ischemia and collagen diseases represent significant problems with few solutions. Cell-based therapy may hold promise in overcoming such disorders. In order to establish a suitable experimental model for the treatment of such ulcers using stem cells, this chapter describes detailed methods for: (1) isolation of stem cells from adipose tissue, termed adipose-derived stem cells (ASCs), (2) preparing a hybrid-type artificial dermis that consists of a type I collagen sponge and ASCs, (3) preparing intractable ulcers using Mitomycin C, and (4) evaluating the effect of wound healing histologically. ASCs seeded onto a type I collagen sponge are applied to intractable ulcers induced by topical application of Mitomycin C. Histological evaluation after 1 and 2 weeks revealed an increase in capillary density and granulation thickness of the hybrid-type artificial dermis. These findings suggest that ASCs may have a positive effect on wound healing and may be a useful tool for future cell-based therapy.
