Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
In Vitro Cell Dev Biol Anim. 2010 Feb;46(2):102-6. doi: 10.1007/s11626-009-9258-6. Epub 2009 Nov 14.
We describe a method for creating differentiated equine bronchial epithelial cell cultures that can be used for in vitro studies including airway disease mechanisms and pathogen-host interactions. Our method is based on the culturing of human tracheobronchial epithelial cells at an air-liquid interface (ALI) in specific serum-free, hormone-supplemented medium. Bronchial epithelial cells are isolated and grown on T-Clear® insert membranes. Within 2 to 3 wk, cells differentiate into ciliated and mucus producing cells as demonstrated by confocal and electron microscopy. Furthermore, the demonstration of the two major gel-forming mucin species, Muc5ac and Muc5b, in our bronchial epithelial cell culture system validates this method for studies of respiratory tract disease of the horse.
我们描述了一种创建分化的马支气管上皮细胞培养物的方法，该方法可用于体外研究，包括气道疾病机制和病原体-宿主相互作用。我们的方法基于在特定的无血清、激素补充培养基中在气液界面 (ALI) 培养人气管支气管上皮细胞。支气管上皮细胞分离并在 T-Clear®插入膜上生长。在 2 到 3 周内，细胞分化为纤毛细胞和分泌粘液的细胞，这一点通过共聚焦和电子显微镜得到证实。此外，我们的支气管上皮细胞培养系统中两种主要的凝胶形成粘蛋白 Muc5ac 和 Muc5b 的证明验证了该方法可用于研究马的呼吸道疾病。
