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

doi:10.1016/j.xpro.2024.103520

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

STAR Protoc. 2025 Jan 7;6(1):103520. doi: 10.1016/j.xpro.2024.103520. Online ahead of print.

Authors

Affiliations

¹ Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada. Electronic address: [email protected].
² Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada.
³ Department of Otolaryngology-Head and Neck Surgery, St. Joseph's Health Care, London, ON, Canada.
⁴ Environmental and Occupational Health, Milken Institute School of Public Health, George Washington University, Washington, D.C., USA.
⁵ Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada. Electronic address: [email protected].

PMID: 39772385
DOI: 10.1016/j.xpro.2024.103520

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.

Keywords: cell culture; cell isolation; microbiology; microscopy.