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

doi:10.1016/j.xpro.2022.101383

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

STAR Protoc. 2022 Apr 22;3(2):101383. doi: 10.1016/j.xpro.2022.101383. eCollection 2022 Jun 17.

Authors

Anne Zebitz Eriksen^{1

2

3

4}, Rasmus Møller⁵, Bar Makovoz^{1

2

3}, Benjamin R tenOever⁵, Timothy A Blenkinsop^{1

2

3}

Affiliations

¹ Department of Cell, Development and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
² Black Family Stem Cell Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
³ Department of Ophthalmology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁴ Department of Health Technology, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
⁵ Department of Microbiology, New York University, New York, NY 10016, USA.

Abstract

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).

Keywords: Health Sciences; Immunology; Microbiology; Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Adult
COVID-19*
Eye
Humans
Organoids
SARS-CoV-2

Abstract

Publication types

MeSH terms

Grants and funding