腺病毒绿色荧光表达载体追踪皮肤组织工程种子细胞的转归  

Tracing seed cells of skin tissue engineering with adenoviral vector harboring green fluorescent protein reporter in vitro

作　　者：刘志国[1] 夏照帆[1] 贲道锋[1] 程大胜[1] 李刚[1] 肖仕初[1] 

机构地区：[1]第二军医大学长海医院烧伤科,上海200433

出　　处：《第二军医大学学报》2002年第12期1325-1327,I003,共4页Academic Journal of Second Military Medical University

基　　金：国家杰出青年科学基金资助项目(3 972 5 0 2 9) ;上海市卫生系统百名跨世纪优秀学科带头人培养计划资助项目 (97BR0 46) ;国家 973项目子课题(G19990 5 43 0 0 -3 ) .

摘　　要：目的 :用转基因技术标记组织工程种子细胞 ,探索一种追踪种子细胞转归的方法。 方法 :用腺病毒荧光表达载体转染人皮肤成纤维细胞 ,观察转染率及其荧光表达。再将细胞种植在真皮支架上 ,活体观察荧光标记的效果和持续时间。结果 :所制备的腺病毒荧光表达载体能转染靶细胞 ,转染率 >95 %。转染的细胞经一次传代后 ,尽管荧光强度降低 ,细胞轮廓仍清晰可见。细胞种植至真皮支架上 ,观察 2周 ,可以在荧光显微镜下较清晰地观察到培养物活体上细胞的形态。结论 :应用腺病毒荧光表达载体标记皮肤组织工程种子细胞以追踪细胞的转归是方便。Objective: To establish a method for tracing seed cells in skin tissue engineering with technology of transgene.Methods: Human skin fibroblasts were transfected with adenoviral vector harboring green fluorescent protein reporter and the transfection rate and the fluorescent protein expression were observed.Transfected cells were seeded on artificial dermis scaffold to observe the labeling effects and its lasting time.Results: The adenoviral vector transfected target cells efficiently and the transfection rate was more than 95%.When transfected cells were subcultured for one time and reached confluence,the profiles of the cells were observed clearly although its fluorescence decreased.Cells in the artificial dermis scaffold were cultured for 2 weeks,the morphology of the cells could be observed clearly in ongoing culture dermis substitute.Conclusion: It is convenient and feasible to trace seed cells of skin tissue engineering with adenoviral vector harboring green fluorescent protein reporter in vitro .

关 键 词：腺病毒绿色荧光表达载体 转基因 组织工程 人工皮肤 种子细胞 成纤维细胞 

分 类 号：R318.1[医药卫生—生物医学工程]