Stead Family Department of Pediatrics, Carver College of Medicine, The University of Iowa, Iowa City, IA, USA.
Scripps Research Institute, University of Florida, Jupiter, FL, USA.
Methods Mol Biol. 2024;2808:141-152. doi: 10.1007/978-1-0716-3870-5_11.
Measles virus (MeV) infection of airway surface epithelial cells provides a site for final amplification before being released back into the environment via coughing and sneezing. Multiple cell lines have served as models of polarized epithelia for MeV infection, such as Caco2 cells (intestinal derived human epithelia) or MDCK cells (kidney derived canine epithelia). In this chapter, we describe the materials and air-liquid interface (ALI) culture conditions for maintaining four different cell lines derived from human airway epithelial cells: 16HBE14o-, Calu-3, H358, and NuLi-1. We provide methods for confirming transepithelial electrical resistance (TER) and preparing samples for microscopy as well as expected results from apical or basolateral MeV delivery. Polarized human airway derived cells serve as tissue culture models for investigating targeted questions about how MeV exits a human host. In addition, these methods are generalizable to studies of other respiratory viruses or the biology of ALI airway epithelial cells.
麻疹病毒（MeV）感染气道表面上皮细胞，为通过咳嗽和打喷嚏将病毒释放回环境中之前提供了最终扩增的场所。多种细胞系已被用作麻疹病毒感染的极化上皮模型，例如 Caco2 细胞（肠道来源的人上皮细胞）或 MDCK 细胞（肾脏来源的犬上皮细胞）。在本章中，我们描述了维持源自人气道上皮细胞的四种不同细胞系（16HBE14o-、Calu-3、H358 和 NuLi-1）的材料和气液界面（ALI）培养条件。我们提供了用于确认跨上皮电阻（TER）和为显微镜准备样品的方法，以及从顶端或基底外侧给予 MeV 的预期结果。极化的人气道衍生细胞作为组织培养模型，用于研究有关 MeV 如何从人体宿主中逸出的靶向问题。此外，这些方法可推广用于研究其他呼吸道病毒或 ALI 气道上皮细胞的生物学。
Zotero 插件