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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| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724006853 |
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| _version_ | 1841553828124557312 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Victor H.K. Lam |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-7c2595f53ce54cc3a60d095b7c2d024e |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-7c2595f53ce54cc3a60d095b7c2d024e2025-01-09T06:14:38ZengElsevierSTAR Protocols2666-16672025-03-0161103520Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory virusesVictor H.K. Lam0Aleena Ghafoor1Yazan Khan2Shirley Constable3Lane B. Buchanan4David Zuanazzi5Reeya Parmar6Zeynep G. Tepe7Leigh J Sowerby8Cindy M. Liu9Ryan M. Troyer10Jessica L. Prodger11Department of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada; Corresponding authorDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Otolaryngology—Head and Neck Surgery, St. Joseph’s Health Care, London, ON, CanadaEnvironmental and Occupational Health, Milken Institute School of Public Health, George Washington University, Washington, D.C., USADepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, CanadaDepartment of Microbiology and Immunology, Schulich School of Medicine & Dentistry, Western University, London, ON, Canada; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166724006853cell culturecell isolationmicrobiologymicroscopy |
| spellingShingle | 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 Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses STAR Protocols cell culture cell isolation microbiology microscopy |
| title | Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses |
| title_full | Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses |
| title_fullStr | Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses |
| title_full_unstemmed | Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses |
| title_short | Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses |
| title_sort | protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses |
| topic | cell culture cell isolation microbiology microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724006853 |
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