中文摘要：

【目的】探讨胚胎来源表皮干细胞构建的人工皮肤模型InspectskinⅠ修复全层皮肤缺损的作用,为构建含皮肤附属结构的检验用皮肤奠定基础。【方法】将人羊膜和BMP-4定向诱导小鼠胚胎细胞分化为表皮干细胞,采用筏式培养在体外与PHBV和成纤维细胞共培养构建组织工程皮肤,经短暂培养后移植于裸鼠皮肤缺损创面,对移植物进行形态学和免疫组织化学观察。【结果】移植物可封闭皮肤缺损,在其表面可形成复层表皮样结构,真皮部分可见大小不等的由单层或复层上皮样细胞构成的管状和泡状结构、汗腺样结构和毛囊样结构。免疫组化和免疫双标结果表明新生表皮的基底层分别呈CK19和CK10阳性,腺管样结构分别呈CEA和CK18阳性。【结论】体外重建的人工皮肤能修复裸鼠全层皮肤缺损,胚胎来源表皮干细胞具有分化为复层扁平上皮、汗腺样结构和毛囊样结构的潜能。

英文摘要：

[Objective] To investigate the effect of restoration of full-thickness skin defect by artificial skin model Inspectskin Ⅰ constructed with embryonic stem （ES） cell-derived epidermal stem cells, and to lay a foundation for constructing new inspective skin model containing appendage. [Methods] Mouse ES cells, which were induced directionally by human amniotic membrane and bone morphogenetic protein-4 （BMP-4）, differentiated into epidermal stem cells which were cocultured with poly（3-hydroxybutyric acid- co-3-hydroxyvaleric acid） （PHBV） and fibroblasts through raft culture in vitro to construct tissue engineering skin. After short culture, the engineering skin was transplanted into the surface of full-thickness skin defect in nude mice. The morphology and immunohistochemistry of donor grafts were observed. [Results] The donor grafts could seal the skin defect, on whose surface stratified epidermal-like structures could be observed. In the dennis, tubular, vesicular, hidradenoid, and hair follicle-like structures constructed by simple epithelium-like or stratified epithelium-like cells with different sizes could be observed. The results of immunohistochemical and di-labeled immunofluorescence methods showed that the basal layers of the new epidermis were cytokeration （CK） 19 and CK10 positive, and the tubular structures were carcinoembryonic antigen （CEA） and CK18 positive. [Conclusions] The artificial skin constructed in vitro can restore the full-thickness skin defect in nude mice and ES-derived epidermal stem cells have the differentiation potential to form stratified squamous epithelium, hidradenoid and hair follicle-like structures.