Mizuno Hiroshi, Nambu Masaki
Department of Plastic and Reconstructive Surgery, School of Medicine, Juntendo University, Tokyo, Japan.
Methods Mol Biol. 2011;702:453-9. doi: 10.1007/978-1-61737-960-4_33.
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.
由糖尿病、缺血和胶原疾病导致的顽固性皮肤溃疡是重大难题,解决方法寥寥无几。基于细胞的疗法可能有望攻克此类病症。为了建立一个使用干细胞治疗此类溃疡的合适实验模型,本章详细描述了以下方法:(1) 从脂肪组织中分离干细胞,即脂肪来源干细胞(ASCs);(2) 制备由I型胶原海绵和ASCs组成的混合型人工真皮;(3) 使用丝裂霉素C制备顽固性溃疡;(4) 从组织学角度评估伤口愈合效果。将接种在I型胶原海绵上的ASCs应用于通过局部涂抹丝裂霉素C诱导产生的顽固性溃疡。1周和2周后的组织学评估显示,混合型人工真皮的毛细血管密度和肉芽厚度增加。这些发现表明,ASCs可能对伤口愈合有积极作用,并且可能成为未来基于细胞疗法的有用工具。