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

Authors

• Victor H. Lam, Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada. Electronic address: hlam242@uwo.ca.
• 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. Electronic address: jprodge@uwo.ca.

Document Type

Journal Article

Publication Date

1-7-2025

Journal

STAR protocols

Volume

6

Issue

1

DOI

10.1016/j.xpro.2024.103520

Keywords

cell culture; cell isolation; microbiology; microscopy

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.

APA Citation

Lam, Victor H.; 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.; and Prodger, Jessica L., "Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses" (2025). GW Authored Works. Paper 6405.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/6405

Department

Environmental and Occupational Health