STAR Protocols, Dec 2022

Elena GarretaDaniel Moya-RullMegan L StaniferVanessa MonteilPatricia PradoAndrés MarcoCarolina TarantinoMaria GalloGustav JonssonAstrid HagelkruysAli MirazimiSteeve Boulant, Josef M PenningerNuria Montserrat

Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells

This protocol presents the use of SARS-CoV-2 isolates to infect human kidney organoids, enabling exploration of the impact of SARS-CoV-2 infection in a human multicellular in vitro system. We detail steps to generate kidney organoids from human pluripotent stem cells (hPSCs) and emulate a diabetic milieu via organoids exposure to diabetogenic-like cell culture conditions. We further describe preparation and titration steps of SARS-CoV-2 virus stocks, their subsequent use to infect the kidney organoids, and assessment of the infection via immunofluorescence. For complete details on the use and execution of this protocol, please refer to Garreta et al. (2022)

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