不同基质对兔气管纤毛上皮细胞贴壁生长的影响  

Effects of Different Substrata on Explant Culture of Rabbit Tracheal Epithelium Ciliated Cells

作　　者：王鸿[1] 张伟[1] 矫健[1] 金善哲[1] 张罗[1,2] 

机构地区：[1]北京市耳鼻咽喉科研究所耳鼻咽喉头颈科学教育部重点实验室 [2]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科

出　　处：《首都医科大学学报》2010年第5期604-609,共6页Journal of Capital Medical University

基　　金：国家自然科学基金(30973282);北京市自然科学基金(7072017);北京市教委科技发展计划项目(KZ200910025008)~~

摘　　要：目的比较几种不同培养基质对兔气管纤毛上皮细胞贴壁生长的影响,探讨兔气管纤毛上皮细胞体外培养最佳生长基质。方法用组织块法比较兔气管纤毛上皮细胞分别在新鲜配制鼠尾胶原、市售鼠尾胶原、多聚赖氨酸和明胶4种基质上的贴壁和生长状况。结果兔气管黏膜上皮组织在4种基质上的贴壁率分别为:新鲜配制胶原89.1%,市售胶原43.5%,多聚赖氨酸36.9%,明胶21.7%;贴壁组织的单细胞层最大生长半径分别为:新鲜配制胶原(1 432.2±383.2)μm,市售胶原(928.9±390.3)μm,多聚赖氨酸(889±229.2)μm,明胶(651±221.2)μm;纤毛摆动最大频率及持续时间分别为:新鲜配制胶原(10.0±1.5)Hz 11 d,市售胶原(8.5±0.6)Hz 5 d,多聚赖氨酸(8.9±0.5)Hz 3 d,明胶(5.9±0.3)Hz 2 d。扫描电镜下新鲜配制鼠尾胶原纤维成条索状,相互交织成网,利于组织贴壁和细胞生长。结论在常用的4种培养基质中,新鲜配制的鼠尾胶原是兔气管纤毛上皮细胞体外培养的最佳生长基质,是纤毛功能研究的可靠保证。Objective To compare the effects of four substrata in cell culture of rabbit tracheal epithelium ciliated cells and to explore the more suitable substratum for the cell model established in vitro.Methods Self-made rat tail collagen,commercial rat tail collagen,poly-L-lysine and gelatin were coated on glass coverslip respectively for explants culture of rabbit tracheal mucosal specimens.Results The anchorage rates of rabbit tracheal mucosal specimens on the four substrata respectively are： self-made rat tail collagen 89.1%,commercial rat tail collagen 43.5%,poly-L-lysine 36.9% and gelatin 21.7%.The maximal monolayer radius of anchorage tissues on four substrata respectively are： self-made rat tail collagen（1 432.2±383.2）μm,commercial rat tail collagen（928.9±390.3）μm,poly-L-lysine（889±229.2）μm and gelatin（651±221.2）μm.The maximal ciliary beat frequency and duration respectively are： self-made rat tail collagen（10.0±1.5）Hz 11 days,commercial rat tail collagen（8.5±0.6）Hz 5 days,poly-L-lysine（8.9±0.5）Hz 3 days and gelatin（5.9±0.3）Hz 2 days.Under the electron microscope,self-made rat tail collagen fibers show a reticulate pattern,which may play an active role in ciliary cell anchorage and growth.Conclusion Our study indicates that self-made rat tail collagen is the most suitable substratum among the four in explants culture of rabbit tracheal epithelium ciliated cells in vitro and could be a reliable cell-attracting substance for ciliary function study.

关 键 词：细胞外基质 纤毛 上皮细胞 鼠尾胶原 

分 类 号：R765[医药卫生—耳鼻咽喉科]