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