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Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading

Our knowledge about respiratory virus spreading is mostly based on monolayer cultures that hardly reflect the complex organization of the airway epithelium. Thus, there is a strong demand for biologically relevant models. One possibility to study virus spreading at the cellular level is real-time im...

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Autores principales: Möckel, Marion, Baldok, Nino, Walles, Thorsten, Hartig, Roland, Müller, Andreas J., Reichl, Udo, Genzel, Yvonne, Walles, Heike, Wiese-Rischke, Cornelia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9688616/
https://www.ncbi.nlm.nih.gov/pubmed/36429061
http://dx.doi.org/10.3390/cells11223634
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author Möckel, Marion
Baldok, Nino
Walles, Thorsten
Hartig, Roland
Müller, Andreas J.
Reichl, Udo
Genzel, Yvonne
Walles, Heike
Wiese-Rischke, Cornelia
author_facet Möckel, Marion
Baldok, Nino
Walles, Thorsten
Hartig, Roland
Müller, Andreas J.
Reichl, Udo
Genzel, Yvonne
Walles, Heike
Wiese-Rischke, Cornelia
author_sort Möckel, Marion
collection PubMed
description Our knowledge about respiratory virus spreading is mostly based on monolayer cultures that hardly reflect the complex organization of the airway epithelium. Thus, there is a strong demand for biologically relevant models. One possibility to study virus spreading at the cellular level is real-time imaging. In an attempt to visualize virus spreading under somewhat more physiological conditions, Calu-3 cells and human primary fibroblasts were co-cultured submerged or as air-liquid interface (ALI). An influenza A virus (IAV) replicating well in cell culture, and carrying a red fluorescent protein (RFP) reporter gene was used for real-time imaging. Our three-dimensional (3D) models exhibited important characteristics of native airway epithelium including a basement membrane, tight junctions and, in ALI models, strong mucus production. In submerged models, first fluorescence signals appeared between 9 and 12 h post infection (hpi) with a low multiplicity of infection of 0.01. Virus spreading further proceeded in the immediate vicinity of infected cells. In ALI models, RFP was found at 22 hpi and later. Consequently, the progression of infection was delayed, in contrast to the submerged model. With these features, we believe that our 3D airway models can deliver new insights in the spreading of IAV and other respiratory viruses.
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spelling pubmed-96886162022-11-25 Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading Möckel, Marion Baldok, Nino Walles, Thorsten Hartig, Roland Müller, Andreas J. Reichl, Udo Genzel, Yvonne Walles, Heike Wiese-Rischke, Cornelia Cells Article Our knowledge about respiratory virus spreading is mostly based on monolayer cultures that hardly reflect the complex organization of the airway epithelium. Thus, there is a strong demand for biologically relevant models. One possibility to study virus spreading at the cellular level is real-time imaging. In an attempt to visualize virus spreading under somewhat more physiological conditions, Calu-3 cells and human primary fibroblasts were co-cultured submerged or as air-liquid interface (ALI). An influenza A virus (IAV) replicating well in cell culture, and carrying a red fluorescent protein (RFP) reporter gene was used for real-time imaging. Our three-dimensional (3D) models exhibited important characteristics of native airway epithelium including a basement membrane, tight junctions and, in ALI models, strong mucus production. In submerged models, first fluorescence signals appeared between 9 and 12 h post infection (hpi) with a low multiplicity of infection of 0.01. Virus spreading further proceeded in the immediate vicinity of infected cells. In ALI models, RFP was found at 22 hpi and later. Consequently, the progression of infection was delayed, in contrast to the submerged model. With these features, we believe that our 3D airway models can deliver new insights in the spreading of IAV and other respiratory viruses. MDPI 2022-11-16 /pmc/articles/PMC9688616/ /pubmed/36429061 http://dx.doi.org/10.3390/cells11223634 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Möckel, Marion
Baldok, Nino
Walles, Thorsten
Hartig, Roland
Müller, Andreas J.
Reichl, Udo
Genzel, Yvonne
Walles, Heike
Wiese-Rischke, Cornelia
Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
title Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
title_full Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
title_fullStr Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
title_full_unstemmed Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
title_short Human 3D Airway Tissue Models for Real-Time Microscopy: Visualizing Respiratory Virus Spreading
title_sort human 3d airway tissue models for real-time microscopy: visualizing respiratory virus spreading
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9688616/
https://www.ncbi.nlm.nih.gov/pubmed/36429061
http://dx.doi.org/10.3390/cells11223634
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