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Methods Mol Biol ; 2273: 131-138, 2021.
Article in English | MEDLINE | ID: covidwho-1092090


The current coronavirus disease-19 (COVID-19) pandemic, caused by "severe acute respiratory syndrome coronavirus 2" (SARS-CoV-2), underscores the threat posed by newly emerging viruses. The understanding of the mechanisms driving early infection events, that are crucial for the exponential spread of the disease, is mandatory and can be significantly implemented generating 3D in vitro models as experimental platforms to investigate the infection substrates and how the virus invades and ravages the tissues.We here describe a protocol for the creation of a synthetic hydrogel-based 3D culture system that mimics in vitro the complex architectures and mechanical cues distinctive of the upper airway epithelia. We then expose the in vitro generated 3D nasal and tracheal epithelia to gold nanoparticles (AuNPs) that display the typical shape and size distinctive of SARS-CoV-2 and of the majority of Coronaviridae presently known.The infection platform here described provides an efficient and highly physiological in vitro model that reproduces the host-pathogen early interactions, using virus-mimicking nanoparticles, and offers a flexible tool to study virus entry into the cell. At the same time, it reduces the risk of accidental infection/spillovers for researchers, which represents a crucial aspect when dealing with a virus that is highly contagious, virulent, and even deadly.

COVID-19/metabolism , COVID-19/virology , Cell Culture Techniques/methods , Epithelial Cells/cytology , Nanoparticles/metabolism , Respiratory Mucosa/cytology , Animals , Cell Line , Chlorocebus aethiops , Epithelial Cells/virology , Gold , Humans , Metal Nanoparticles/chemistry , Molecular Mimicry/immunology , Nose/virology , Respiratory Mucosa/virology , SARS-CoV-2/metabolism , SARS-CoV-2/pathogenicity , Trachea/virology , Vero Cells , Virus Internalization