使用呼吸道病毒成像生成和表征鼻上皮模型的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS STAR Protocols Pub Date : 2025-01-07 DOI:10.1016/j.xpro.2024.103520
Victor H K Lam, Aleena Ghafoor, Yazan Khan, Shirley Constable, Lane B Buchanan, David Zuanazzi, Reeya Parmar, Zeynep G Tepe, Leigh J Sowerby, Cindy M Liu, Ryan M Troyer, Jessica L Prodger
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引用次数: 0

摘要

气液界面(ALI)培养可使气道上皮细胞分化成体外呼吸道。在这里,我们提出了一种分离和培养严重急性呼吸综合征冠状病毒2 (SARS-CoV-2)感染鼻甲组织鼻上皮细胞的方案。我们描述了克服成像脆弱培养物挑战的步骤,检测粘液的产生,并量化sars - cov -2感染后的细胞内病毒。我们在实验之前提供了关于ALI成熟的最佳持续时间的数据,并描述了可以改变哪些步骤来优化特定假设的测试。
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Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses.

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.

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来源期刊
STAR Protocols
STAR Protocols Biochemistry, Genetics and Molecular Biology-General Biochemistry, Genetics and Molecular Biology
CiteScore
2.00
自引率
0.00%
发文量
789
审稿时长
10 weeks
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