Protocols for SARS-CoV-2 infection in primary ocular cells and eye organoids

Here, we describe a series of protocols detailing the steps for evaluating SARS-CoV-2 infection in models of the human eye. Included are protocols for whole eye organoid differentiation, SARS-CoV-2 infection, and processing organoids for single-cell RNA sequencing. Additional protocols describe how...

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Detalles Bibliográficos
Autores principales: Eriksen, Anne Zebitz, Møller, Rasmus, Makovoz, Bar, tenOever, Benjamin R., Blenkinsop, Timothy A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9023348/
https://www.ncbi.nlm.nih.gov/pubmed/35664254
http://dx.doi.org/10.1016/j.xpro.2022.101383
Descripción
Sumario:Here, we describe a series of protocols detailing the steps for evaluating SARS-CoV-2 infection in models of the human eye. Included are protocols for whole eye organoid differentiation, SARS-CoV-2 infection, and processing organoids for single-cell RNA sequencing. Additional protocols describe how to dissect and culture adult human ocular cells from cadaver donor eyes and how to compare infection of SARS-CoV-2 and the presence of SARS-CoV-2 entry factors using qPCR, immunofluorescence, and plaque assays. For complete details on the use and execution of this protocol, please refer to Eriksen et al. (2021).

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