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Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2.

STAR protocols (2022-08-23)
Hyunjae Chung, Maxwell P Bui-Marinos, Waleed Rahmani, Jennifer A Corcoran, Justin Chun
RÉSUMÉ

Induced pluripotent stem cell (iPSC)-derived kidney organoids can be used for disease modeling and drug testing. Here, we describe a protocol to prepare stocks of an infectious clone of SARS-CoV-2 expressing a stable mNeonGreen reporter (icSARS-CoV-2-mNG). We demonstrate the infection of kidney organoids, primarily at the proximal tubular cells, with icSARS-CoV-2-mNG. Using a TCID50 (tissue culture infectious dose 50) assay and confocal microscopy, we show the quantification of SARS-CoV-2-mNG signal in proximal tubular cells of the kidney organoids. For complete details on the use and execution of this protocol, please refer to Rahmani et al. (2022).

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Sérum de veau fœtal, Canada origin, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Y-27632, InSolution, ≥95%, reversible, inhibitor of Rho kinases