Horani Amjad, Dickinson John D, Brody Steven L
Department of Pediatrics, Division of Pediatric Allergy, Immunology, and Pulmonary Medicine, Washington University in St. Louis, Saint Louis, MO, USA.
Methods Mol Biol. 2013;1032:91-107. doi: 10.1007/978-1-62703-496-8_7.
Primary airway epithelial cell culture provides a valuable tool for studying cell differentiation, cell-cell interactions, and the role of immune system factors in asthma pathogenesis. In this chapter, we discuss the application of mouse tracheal epithelial cell cultures for the study of asthma biology. A major advantage of this system is the ability to use airway epithelial cells from mice with defined genetic backgrounds. The in vitro proliferation and differentiation of mouse airway epithelial cells uses the air-liquid interface condition to generate well-differentiated epithelia with characteristics of native airways. Protocols are provided for manipulation of differentiation, induction of mucous cell metaplasia, genetic modification, and cell and pathogen coculture. Assays for the assessment of gene expression, responses of cells, and analysis of specific cell subpopulations within the airway epithelium are included.
原代气道上皮细胞培养为研究细胞分化、细胞间相互作用以及免疫系统因子在哮喘发病机制中的作用提供了一种有价值的工具。在本章中,我们将讨论小鼠气管上皮细胞培养在哮喘生物学研究中的应用。该系统的一个主要优点是能够使用具有明确遗传背景的小鼠的气道上皮细胞。小鼠气道上皮细胞的体外增殖和分化利用气液界面条件来生成具有天然气道特征的高度分化上皮。文中提供了用于调控分化、诱导黏液细胞化生、基因修饰以及细胞与病原体共培养的方案。还包括用于评估基因表达、细胞反应以及分析气道上皮内特定细胞亚群的检测方法。