利用成像技术生成和表征用于呼吸道病毒研究的鼻上皮模型的方案

Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses.

作者信息

Lam Victor H K, Ghafoor Aleena, Khan Yazan, Constable Shirley, Buchanan Lane B, Zuanazzi David, Parmar Reeya, Tepe Zeynep G, Sowerby Leigh J, Liu Cindy M, Troyer Ryan M, Prodger Jessica L

机构信息

Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103520. doi: 10.1016/j.xpro.2024.103520. Epub 2025 Jan 7.

DOI:10.1016/j.xpro.2024.103520

PMID:39772385

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11760824/

Abstract

Air-liquid interface (ALI) culture can differentiate airway epithelial cells to recapitulate the respiratory tract in vitro. Here, we present a protocol for isolating and culturing nasal epithelial cells from turbinate tissues for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. We describe steps to overcome challenges of imaging fragile cultures, detect the production of mucus, and quantify intracellular virus post-SARS-CoV-2 infection. We present data on the optimal duration of ALI maturation prior to experimentation and describe which steps can be altered to optimize testing of specific hypotheses.

摘要

气液界面（ALI）培养可使气道上皮细胞分化，从而在体外重现呼吸道。在此，我们展示了一种从鼻甲组织中分离和培养鼻上皮细胞以用于严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染的方案。我们描述了克服对脆弱培养物进行成像的挑战、检测黏液产生以及量化SARS-CoV-2感染后细胞内病毒的步骤。我们展示了实验前ALI成熟的最佳持续时间的数据，并描述了哪些步骤可以改变以优化对特定假设的测试。