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

Elena Garreta; Daniel Moya-Rull; Megan L. Stanifer; Vanessa Monteil; Patricia Prado; Andrés Marco; Carolina Tarantino; Maria Gallo; Gustav Jonsson; Astrid Hagelkruys; Ali Mirazimi; Steeve Boulant; Josef M. Penninger; Nuria Montserrat

STAR Protocols (Dec 2022)

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

Elena Garreta,
Daniel Moya-Rull,
Megan L. Stanifer,
Vanessa Monteil,
Patricia Prado,
Andrés Marco,
Carolina Tarantino,
Maria Gallo,
Gustav Jonsson,
Astrid Hagelkruys,
Ali Mirazimi,
Steeve Boulant,
Josef M. Penninger,
Nuria Montserrat

Affiliations

Elena Garreta: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
Daniel Moya-Rull: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
Megan L. Stanifer: Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany; Research Group “Cellular Polarity and Viral Infection”, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, USA
Vanessa Monteil: Karolinska Institute and Karolinska University Hospital, Unit of Clinical Microbiology, 17182 Stockholm, Sweden
Patricia Prado: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
Andrés Marco: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
Carolina Tarantino: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
Maria Gallo: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
Gustav Jonsson: IMBA, Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Dr. Bohr-Gasse 3, 1030 Vienna, Austria
Astrid Hagelkruys: IMBA, Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Dr. Bohr-Gasse 3, 1030 Vienna, Austria
Ali Mirazimi: Karolinska Institute and Karolinska University Hospital, Unit of Clinical Microbiology, 17182 Stockholm, Sweden; National Veterinary Institute, 751 89 Uppsala, Sweden; Corresponding author
Steeve Boulant: Research Group “Cellular Polarity and Viral Infection”, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, USA; Department of Infectious Diseases, Virology, Heidelberg University Hospital, Heidelberg, Germany; Corresponding author
Josef M. Penninger: IMBA, Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Dr. Bohr-Gasse 3, 1030 Vienna, Austria; Department of Medical Genetics, Life Sciences Institute, University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Corresponding author
Nuria Montserrat: Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain; Catalan Institution for Research and Advanced Studies (ICREA), Barcelona, Spain; Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina, 28029 Madrid, Spain; Corresponding author

Journal volume & issue: Vol. 3, no. 4
p. 101872

Abstract

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Summary: 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).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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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