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

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

STAR Protocols (Mar 2025)

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

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

Affiliations

Victor H.K. Lam: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada; Corresponding author
Aleena Ghafoor: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Yazan Khan: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Shirley Constable: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Lane B. Buchanan: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
David Zuanazzi: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Reeya Parmar: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Zeynep G. Tepe: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Leigh J Sowerby: Department of Otolaryngology—Head and Neck Surgery, St. Joseph’s Health Care, London, ON, Canada
Cindy M. Liu: Environmental and Occupational Health, Milken Institute School of Public Health, George Washington University, Washington, D.C., USA
Ryan M. Troyer: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada
Jessica L. Prodger: Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada; Corresponding author

Journal volume & issue: Vol. 6, no. 1
p. 103520

Abstract

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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.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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