小鼠原代呼吸道上皮细胞分离培养的新方法  

A new method for isolation and culture of tracheal epithelial cell of mice

作　　者：王达[1] 李煜生[2] 

机构地区：[1]南方医科大学第一临床医学院,广州510515 [2]南方医科大学基础医学院病理生理学教研室广东省蛋白质组学重点实验室和教育部重大疾病的转录组和蛋白质组学重点实验室,广州510515

出　　处：《生理学报》2011年第6期581-585,共5页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China for Youth(No.81100008);the Foundation for Distinguished Young Talents in Higher Education of Guangdong Province;China(No.LYM10048)

摘　　要：现有的呼吸道上皮细胞系大多来源于肿瘤组织或成系时与肿瘤细胞融合,其生物学行为与正常呼吸道上皮细胞差异较大。为更准确反映呼吸道疾病条件下该类细胞的生物学效应,本文对小鼠呼吸道上皮细胞分离的新技术和培养方法进行了探索。利用链霉蛋白酶消化法分离获得小鼠呼吸道上皮细胞,利用特殊的完全培养基和I型胶原包被的培养皿进行原代培养。镜下可观察到呼吸道上皮细胞的纤毛节律性的定向摆动,证明本法所获得的细胞较完整地保留了其原有生理功能。本实验所建立的分离技术和培养方法简单、稳定、有效、可靠,为研究呼吸道疾病的细胞模型创造了条件。同时,此法可为其它实验动物呼吸道上皮细胞的培养提供借鉴。Since most of the respiratory epithelial cell lines are descended from neoplastic tissues or have been fused with neoplastic cells when being selected to a cell line, their biological behaviors are far different from normal respiratory epithelial cells. Aiming at better reflecting the biological properties of epithelial cells under respiratory pathological conditions, our study probed into a new iso- lation technique and culture method of mice respiratory epithelial cell. Pronase was applied for cell isolation from mouse trachea, and a modified medium with collagen-coated plate for primary culture. The ciliary movement can be observed under microscope, which demonstrates that the original biological functions of the tracheal epithelial cell have been reserved with our method. The research presents the efficacy, convenience and reliability of the separation technique and culture method established by this study, and laying preferable condition for cell models of respiratory diseases. Meanwhile, this method may be used for other animal models.

关 键 词：呼吸道上皮细胞 小鼠 分离 培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]