兔气管黏膜上皮细胞片的制备  

The creation of rabbit tissue engineered tracheal epithelial cell sheets

作　　者：张卫东 章方彪 史宏灿 韩石 叶钢 孙飞 潘枢 刘星辰 

机构地区：[1]扬州大学临床医学院扬州大学转化医学研究中心,225001

出　　处：《中华实验外科杂志》2014年第11期2462-2464,I0001,I0002,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81170014、81370118）

摘　　要：目的 探讨兔气管黏膜上皮细胞片的体外制备方法.方法 使用组织块法体外分离培养兔气管黏膜上皮细胞,进行鉴定后,将其种植于UpCell培养皿,待细胞长满皿底后,将环境温度降至20℃,利用培养皿温敏特性制备出上皮细胞片.结果 利用组织块法分离培养的兔气管黏膜上皮细胞,角蛋白CK-18表达阳性,经过UpCell培养皿的培养,当降低温度至20℃时,可分离出完整的上皮细胞片,经鉴定细胞片内的细胞连接紧密,并相互叠加.结论 利用UpCell培养皿的温敏反应性可成功的制备出完整上皮细胞片,上皮细胞片为组织工程气管的构建提供了新的策略.Objective To establish a method for creating rabbit tracheal epithelial cell sheets in vitro.Methods Rabbit tracheal epithelial cells were isolated and cultured using tissue explant technique,and passaged in UpCell culture dishes at 37 ℃,5% CO2 and saturated humidity after being identified by cytokeratin （CK）-18 staining.About two days,cells have reached confluency,and tracheal epithelial cell sheet was prepared when the ambient temperature was changed to 20 ℃.Results Epithelial cells were isolated from rabbit tracheal mucosa using tissue explant technique successfully,and all cells were positive for CK-18 staining,showing they had properties of epithelial cells.When the ambient temperature dropped at 20 ℃,epithelial cells detached automatically from the UpCell culture dishes and formed an intact cell sheet.The cells are strongly attached and overlapping under light microscope.Conclusion Rabbit tracheal epithelial cell sheet can be fabricated with UpCell culture dishes and it could provides a novel pathway for constructing tissue engineering trachea.

关 键 词：兔 上皮细胞 细胞片 组织工程气管 UpCell培养皿 

分 类 号：R361.2[医药卫生—病理学]