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

Anne Zebitz Eriksen; Rasmus Møller; Bar Makovoz; Benjamin R. tenOever; Timothy A. Blenkinsop

STAR Protocols (Jun 2022)

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

Anne Zebitz Eriksen,
Rasmus Møller,
Bar Makovoz,
Benjamin R. tenOever,
Timothy A. Blenkinsop

Affiliations

Anne Zebitz Eriksen: Department of Cell, Development and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Ophthalmology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Health Technology, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark; Corresponding author
Rasmus Møller: Department of Microbiology, New York University, New York, NY 10016, USA
Bar Makovoz: Department of Cell, Development and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Ophthalmology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
Benjamin R. tenOever: Department of Microbiology, New York University, New York, NY 10016, USA
Timothy A. Blenkinsop: Department of Cell, Development and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Black Family Stem Cell Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Ophthalmology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101383

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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