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

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

Summary: 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)...

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Bibliographic Details
Main 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
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:STAR Protocols
Subjects:
cell culture
cell isolation
microbiology
microscopy
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724006853
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Summary:Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667

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