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Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
Marion Möckel1, Nino Baldok1,2, Thorsten Walles1
1University Clinic for Cardiac and Thoracic Surgery, Otto-von-Guericke-University Magdeburg, D-39120 Magdeburg, Germany.
Cells
|November 26, 2022
Summary
New 3D airway models using co-cultured cells offer a more accurate view of respiratory virus spread. These biologically relevant models visualize influenza A virus (IAV) infection dynamics in real-time, revealing differences between submerged and air-liquid interface cultures.
Area of Science:
- * Virology
- * Cell Biology
- * Respiratory Medicine
Background:
- * Current understanding of respiratory virus spread relies on simplistic monolayer cell cultures.
- * These models fail to replicate the complex structure and function of native airway epithelium.
- * There is a critical need for more biologically relevant models to study virus-host interactions.
Purpose of the Study:
- * To develop and characterize novel three-dimensional (3D) airway models for studying respiratory virus dissemination.
- * To visualize the real-time spread of influenza A virus (IAV) in these advanced models.
- * To compare virus spreading dynamics in submerged versus air-liquid interface (ALI) culture conditions.
Main Methods:
- * Co-culture of Calu-3 cells and human primary fibroblasts in submerged and ALI configurations.
- * Infection with a red fluorescent protein (RFP)-expressing influenza A virus (IAV).
- * Real-time imaging to monitor virus replication and spread.
Main Results:
- * The 3D models successfully recapitulated key features of native airway epithelium, including basement membranes, tight junctions, and mucus production (in ALI models).
- * In submerged models, initial influenza A virus (IAV) fluorescence was observed at 9-12 hours post-infection (hpi) with rapid local spread.
- * In ALI models, fluorescence appeared later (≥22 hpi), indicating a delayed infection progression compared to submerged cultures.
Conclusions:
- * Developed 3D airway models provide a more physiologically relevant platform for studying respiratory virus spread.
- * These models enable real-time visualization of virus dynamics under conditions mimicking native airway epithelium.
- * The findings highlight the potential of these 3D models to yield new insights into the pathogenesis of influenza A virus (IAV) and other respiratory viruses.
