【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。更多还原

【Abstract】 OBJECTIVE To develop an explant outgrowth culture system for airway ciliated 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 Photo- contrast 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.更多还原