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

{"title":"使用呼吸道病毒成像生成和表征鼻上皮模型的方案。","authors":"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","doi":"10.1016/j.xpro.2024.103520","DOIUrl":null,"url":null,"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.","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103520"},"PeriodicalIF":1.3000,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses.\",\"authors\":\"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\",\"doi\":\"10.1016/j.xpro.2024.103520\",\"DOIUrl\":null,\"url\":null,\"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.\",\"PeriodicalId\":34214,\"journal\":{\"name\":\"STAR Protocols\",\"volume\":\"6 1\",\"pages\":\"103520\"},\"PeriodicalIF\":1.3000,\"publicationDate\":\"2025-01-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"STAR Protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1016/j.xpro.2024.103520\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2024.103520","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

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。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

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.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊