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

doi:10.1016/j.xpro.2022.101872

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

STAR Protoc. 2022 Dec 16;3(4):101872. doi: 10.1016/j.xpro.2022.101872. Epub 2022 Nov 7.

Authors

Affiliations

¹ Pluripotency for Organ Regeneration, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain.
² 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.
³ Karolinska Institute and Karolinska University Hospital, Unit of Clinical Microbiology, 17182 Stockholm, Sweden.
⁴ IMBA, Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Dr. Bohr-Gasse 3, 1030 Vienna, Austria.
⁵ Karolinska Institute and Karolinska University Hospital, Unit of Clinical Microbiology, 17182 Stockholm, Sweden; National Veterinary Institute, 751 89 Uppsala, Sweden. Electronic address: ali.mirazimi@ki.se.
⁶ 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. Electronic address: s.boulant@ufl.edu.
⁷ 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. Electronic address: josef.penninger@ubc.ca.
⁸ 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. Electronic address: nmontserrat@ibecbarcelona.eu.

Abstract

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.

Keywords: Cell Differentiation; Cell culture; Microbiology; Microscopy; Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19*
Cell Differentiation
Humans
Kidney
Organoids
Pluripotent Stem Cells*
SARS-CoV-2

Abstract

Publication types

MeSH terms

Grants and funding