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 m...

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Autores principales: Garreta, Elena, Moya-Rull, Daniel, Stanifer, Megan L., Monteil, Vanessa, Prado, Patricia, Marco, Andrés, Tarantino, Carolina, Gallo, Maria, Jonsson, Gustav, Hagelkruys, Astrid, Mirazimi, Ali, Boulant, Steeve, Penninger, Josef M., Montserrat, Nuria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9637521/
https://www.ncbi.nlm.nih.gov/pubmed/36595951
http://dx.doi.org/10.1016/j.xpro.2022.101872
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author Garreta, Elena
Moya-Rull, Daniel
Stanifer, Megan L.
Monteil, Vanessa
Prado, Patricia
Marco, Andrés
Tarantino, Carolina
Gallo, Maria
Jonsson, Gustav
Hagelkruys, Astrid
Mirazimi, Ali
Boulant, Steeve
Penninger, Josef M.
Montserrat, Nuria
author_facet Garreta, Elena
Moya-Rull, Daniel
Stanifer, Megan L.
Monteil, Vanessa
Prado, Patricia
Marco, Andrés
Tarantino, Carolina
Gallo, Maria
Jonsson, Gustav
Hagelkruys, Astrid
Mirazimi, Ali
Boulant, Steeve
Penninger, Josef M.
Montserrat, Nuria
author_sort Garreta, Elena
collection PubMed
description 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)
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spelling pubmed-96375212022-11-07 Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells Garreta, Elena Moya-Rull, Daniel Stanifer, Megan L. Monteil, Vanessa Prado, Patricia Marco, Andrés Tarantino, Carolina Gallo, Maria Jonsson, Gustav Hagelkruys, Astrid Mirazimi, Ali Boulant, Steeve Penninger, Josef M. Montserrat, Nuria STAR Protoc Protocol 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) Elsevier 2022-11-07 /pmc/articles/PMC9637521/ /pubmed/36595951 http://dx.doi.org/10.1016/j.xpro.2022.101872 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Garreta, Elena
Moya-Rull, Daniel
Stanifer, Megan L.
Monteil, Vanessa
Prado, Patricia
Marco, Andrés
Tarantino, Carolina
Gallo, Maria
Jonsson, Gustav
Hagelkruys, Astrid
Mirazimi, Ali
Boulant, Steeve
Penninger, Josef M.
Montserrat, Nuria
Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells
title Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells
title_full Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells
title_fullStr Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells
title_full_unstemmed Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells
title_short Protocol for SARS-CoV-2 infection of kidney organoids derived from human pluripotent stem cells
title_sort protocol for sars-cov-2 infection of kidney organoids derived from human pluripotent stem cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9637521/
https://www.ncbi.nlm.nih.gov/pubmed/36595951
http://dx.doi.org/10.1016/j.xpro.2022.101872
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