呼吸道纤毛上皮细胞的组织块法培养  

An explant outgrowth culture system for airway ciliated cells

作　　者：王奎吉[1] 张罗[1] 韩德民[1] 王鸿[1] 范尔钟[1] 宋晓红[1] 刘仲燕[1] 

机构地区：[1]北京市耳鼻咽喉科研究所,北京100005

出　　处：《中国耳鼻咽喉头颈外科》2006年第12期833-837,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：国家自然科学基金资助项目(30572026);北京市科技新星计划资助项目(2003-B-15);首都医科大学临床基础合作项目(2004ZR08)联合资助

摘　　要：目的建立组织块法呼吸道纤毛上皮细胞原代培养模型,观察细胞生长分化情况,为深入研究黏液纤毛传输系统提供方法学基础。方法应用组织块法在鼠尾胶原上培养兔气管纤毛上皮细胞,对培养7天的标本行苏木素-伊红染色、黏蛋白5AC(mucin-5AC,MUC5AC)单克隆抗体免疫组织化学染色、扫描电镜及透射电镜观察、纤毛摆动频率测量。结果①细胞培养7天后可以见到大量的纤毛细胞和杯状细胞,纤毛形态正常,摆动活跃;②苏木素-伊红染色和黏蛋白MUC5AC单克隆抗体免疫组织化学染色可见分化良好的纤毛细胞和杯状细胞;③扫描电镜及透射电镜观察见纤毛细胞分化良好,纤毛形态正常;④(30±1)℃下测得纤毛摆动频率为(13.2±0.9)次/s。结论应用组织块法进行兔气管纤毛上皮细胞培养,可以得到分化良好的纤毛细胞和杯状细胞,纤毛摆动活跃,可应用此模型对黏液纤毛传输系统进行深入研究。OBJECTIVE To develop an explant outgrowth culture system for airway cJlJated cells and explore the usefulness of the system.METHODS Mucosal specimens from rabbit tracheal epithelium were seeded into collagen-coated cover-slips. After 7 days,the specimens were characterized by Photocontrast microscopy, transmission and Scanning electron microscope. They were also examined with hematoxylin-eosin staining and mucin-5AC immunohistochemistry. Ciliary beat frequency was measured with high-speed digital microscopy. RESULTS Many ciliated cells with good viability and goblet cells were found in the culture system. Immunohistochemistry analysis revealed that mucin- 5AC antibody labeled mainly in the goblet cells.The ciliated cells were differentiated well detected by electron microscope. At （30±1） ℃,the basal ciliary beat frequency was （13.2±0.9） Hz. CONCLUSION The method may be useful in establishing a culture system for ciliated cells and the system will be suitable for the long-term studies of mucociliary transport system.

关 键 词：纤毛 上皮细胞 细胞培养 粘蛋白类 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]