人源性组织工程皮肤白念珠菌感染模型的构建  

Development of a model for Candlda albicans infection using tissue-engineered human skin substitutes

作　　者：曹玉萍[1] 沈永年[1] 吕桂霞[1] 王乐[1] 曾荣[1] 蔡晴[1] 李玲珺[1] 马鹏程[1] 刘维达[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所,南京210042

出　　处：《中华皮肤科杂志》2013年第8期551-553,共3页Chinese Journal of Dermatology

基　　金：国家科技重大专项（2012ZX09301002-005）;973国家重点基础研究发展计划（2013CB531605）;中央级公益性科研院所基本科研业务专项基金（2011PYS006）;江苏省自然科学基金（BK2008092）;江苏省临床医学科技专项（BL2012003）

摘　　要：目的利用人皮肤来源的细胞体外构建含色素组织工程皮肤白念珠菌感染模型。方法利用人皮肤原代培养成纤维细胞、角质形成细胞、黑素细胞，成纤维细胞与胶原孵育3d后得到组织工程真皮，在其表面接种角质形成细胞和黑素细胞培养12d，获得组织工程皮肤，再在其表面滴加1个麦氏浓度的白念珠菌悬液5μl，分别在6、12、24、48和72h进行HE染色和PAS染色观察。结果利用成纤维细胞、角质形成细胞、黑素细胞及胶原构建的组织工程皮肤结构良好，表皮分化层次分明。HE染色和PAS染色显示，感染白念珠菌后，随着时间的延长，白念珠菌对表皮结构的破坏逐渐加重，菌丝由浅表向真皮逐渐侵入。感染至72h，表皮结构完全破坏，真皮内充满大量菌丝。结论利用人皮肤来源的细胞，培养出的含色素组织工程皮肤层次结构分明；初步建立了人皮肤来源的组织工程皮肤念珠菌感染模型。Objective To develop a model for Candida albicans infection using a pigmented tissue- engineered skin substitute derived from human skin cells. Methods Fibroblasts, keratinocytes and melanoeytes were isolated from the foreskin of Chinese adults, and subjected to serial passages. Fibroblasts were incubated in collagen for three days to construct tissue-engineered dermis. Then, keratinocytes and melanoeytes were co- cultured above the dermis for 12 days to construct epidermis. Subsequently, the constructed tissue-engineered skin was inoculated with 5 μl of C. albicans suspension （106 spores/ml） followed by culture for 6, 12, 24, 48 and 72 hours. Hematoxylin-eosin （HE） staining and periodic acid-Schiff （PAS） staining were carried out to observe the infected skin constitute. Results The tissue-engineered skin constructed by fibroblasts, keratinocytes, melanocytes and collagen showed a well-organized structure and highly differentiated epidermis. After infection with C. albicans, the epidermal structure was gradually destructed with the extension of hyphae from the superficial epidermis to dermis, and was completely destroyed with the dermis filled with hyphae at 72 hours. Conclusions Pigmented tissue-engineered skin is successfully constructed using human skin cells with a well-organized structure and highly differentiated epidermis, and a model for C. albicans infection has also been preliminarily established on the pigmented tissue-engineered skin.

关 键 词：组织工程 皮肤 人工 念珠菌 白色 模型 生物学 

分 类 号：R7[医药卫生—临床医学]