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Development of a Vascularized Skin Construct Using Adipose-Derived Stem Cells from Debrided Burned Skin

机译:使用清创性烧伤皮肤中的脂肪干细胞开发血管化皮肤构造物

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Large body surface area burns pose significant therapeutic challenges. Clinically, the extent and depth of burn injury may mandate the use of allograft for temporary wound coverage while autografts are serially harvested from the same donor areas. The paucity of donor sites in patients with burns involving large surface areas highlights the need for better skin substitutes that can achieve early and complete coverage and retain normal skin durability with minimal donor requirements. We have isolated autologous stem cells from the adipose layer of surgically debrided burned skin (dsASCs), using a point-of-care stem cell isolation device. These cells, in a collagen—polyethylene glycol fibrin-based bilayer hydrogel, differentiate into an epithelial layer, a vascularized dermal layer, and a hypodermal layer.All-trans-retinoicacid and fenofibrate were used to differentiate dsASCs into epithelial-like cells. Immunocytochemical analysis showed a matrix- and time-dependent change in the expression of stromal, vascular, and epithelial cell markers. These results indicate that stem cells isolated from debrided skin can be used as a single autologous cell source to develop a vascularized skin construct without culture expansion or addition of exogenous growth factors. This technique may provide an alternative approach for cutaneous coverage after extensive burn injuries.
机译:较大的身体表面积灼伤构成重大的治疗挑战。临床上,烧伤的程度和深度可能要求使用同种异体移植物临时覆盖伤口,而自体移植物是从相同供体区域连续收获的。烧伤患者涉及大表面积的供体部位很少,这凸显了对更好的皮肤替代品的需求,这些替代品可实现早期和完全覆盖,并以最小的供体需求保持正常的皮肤耐久性。我们已经使用即时医疗干细胞分离装置从外科清创烧伤皮肤(dsASCs)的脂肪层中分离出自体干细胞。这些细胞在胶原蛋白-基于聚乙二醇纤维蛋白的双层水凝胶中分化为上皮层,带血管的真皮层和皮下层。全反式维甲酸和非诺贝特用于将dsASCs分化为上皮样细胞。免疫细胞化学分析显示基质,血管和上皮细胞标志物表达的基质和时间依赖性变化。这些结果表明,从清创的皮肤分离的干细胞可以用作单一的自体细胞来源,以发展出血管化的皮肤结构,而无需培养物扩增或添加外源性生长因子。该技术可为广泛的烧伤后的皮肤覆盖提供替代方法。

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