应用表皮干细胞与去表皮的真皮体外构建组织工程皮肤  

Construction of tissue-engineered skin with epidermal stem cells and de-epidermized dermis

作　　者：王新宇[1] 陆洪光[1] 卜晓琳[1] 彭琳琳[1] 赵志国[1] 汤朝武[2] 王洪涛[2] 

机构地区：[1]贵阳医学院附属医院皮肤科,550004 [2]第四军医大学西京医院烧伤科

出　　处：《中华皮肤科杂志》2008年第7期458-461,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30571685）

摘　　要：目的用人皮肤表皮干细胞（ESCs）接种至同种去表皮的真皮上，体外构建组织工程皮肤。方法用两步酶消化法分离获得角质形成细胞，并用Ⅳ型胶原快速粘附法筛选ESCs并培养；进行细胞形态学观察和用免疫组化法对ESCs进行鉴定；将ESCs接种至去表皮的真皮上浸没培养7d后转成气-液面培养，4周后取培养的组织工程皮肤切片分别做HE染色和角蛋白的免疫组化染色。结果培养的ESCs呈铺路石样、克隆状生长；角蛋白K19和β1-整合素的免疫组化染色阳性。培养5周的组织工程皮肤可见有3～6层细胞，并可见角质层，表皮角蛋白染色阳性。结论应用人皮肤的ESCs与去表皮的真皮可以体外构建组织工程皮肤。Objective To construct tissue-engineered skin via in vitro inoculation of epidermal stem cells （ESCs） onto de-epidermized dermis. Methods Skin tissue was obtained from the foreskin of a healthy 6-year-old child, and keratinocytes were isolated by two-step trypsinization method followed by the collection of ESCs via rapid adhesion by collagen Ⅳ. The ESCs were identified by morphological observation and immunohistochemical staining with K19 and integrin β1. To construct tissue-engineered skin, selected ESCs were seeded onto the surface of de-epidermized dermis followed by a one-week culture immersed in the medium and a subsequent 4-week culture at the air-medium interface. The tissue-engineered skin was evaluated with haematoxylin ＆ eosin （HE） staining as well as keratin immunohistochemistry. Results Microscopically, cultured ESCs showed a paving stone-like appearance and grew into colonies. Immunohistochemistry revealed that the ESCs were positive for integrin-β1 and keratin 19. After 5 weeks of culture, 3 - 6 layers of epidermal cell were observed on the dermis with the formation of stratum corneum. Keratin protein was observed in the artificial epidermal skin. Conclusion Tissue-engineered skin is successfully constructed with epidermal stem cells and de-epidermized dermis in vitro.

关 键 词：人工皮肤 干细胞 表皮 真皮 

分 类 号：R318.0[医药卫生—生物医学工程]