Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level.

COVID-19 Humans Induced Pluripotent Stem Cells Kidney Organoids SARS-CoV-2
Cell biology Microbiology Organoids Single cell Stem cells

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
16 09 2022
Historique:
entrez: 19 8 2022
pubmed: 20 8 2022
medline: 23 8 2022
Statut: epublish

Résumé

We describe a protocol for single-cell RNA sequencing of SARS-CoV-2-infected human induced pluripotent stem cell (iPSC)-derived kidney organoids. After inoculation of kidney organoids with virus, we use mechanical and enzymatic disruption to obtain single cell suspensions. Next, we process the organoid-derived cells into sequencing-ready SARS-CoV-2-targeted libraries. Subsequent sequencing analysis reveals changes in kidney cells after virus infection. The protocol was designed for kidney organoids cultured in a 6-well transwell format but can be adapted to organoids with different organ backgrounds. For complete details on the use and execution of this protocol, please refer to Jansen et al. (2022).

Identifiants

DOI: 10.1016/j.xpro.2022.101612 PMID: 35983169 PMC: PMC9293950
pubmed: 35983169
doi: 10.1016/j.xpro.2022.101612
pii: S2666-1667(22)00492-0
pmc: PMC9293950
pii:
doi:

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

101612

Informations de copyright

© 2022 The Author(s).

Déclaration de conflit d'intérêts

The authors declare no competing interests.

Références

Viruses. 2021 Feb 11;13(2):
pubmed: 33670363
Cell Stem Cell. 2022 Feb 3;29(2):217-231.e8
pubmed: 35032430
Mol Syst Biol. 2021 Apr;17(4):e10232
pubmed: 33904651
Nat Methods. 2020 Jun;17(6):629-635
pubmed: 32483332
Nature. 2020 Sep;585(7826):588-590
pubmed: 32698190
Nat Protoc. 2016 Sep;11(9):1681-92
pubmed: 27560173
Cell. 2006 Aug 25;126(4):663-76
pubmed: 16904174

Auteurs

Katharina C Reimer (KC)

Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, Aachen, Germany; Institute of Cell and Tumor Biology, RWTH Aachen University, Aachen, Germany; Division of Nephrology and Clinical Immunology, RWTH Aachen University, Aachen, Germany. Electronic address: katharina.reimer@posteo.de.

Jitske Jansen (J)

Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, Aachen, Germany; Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands; Department of Pediatric Nephrology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Amalia Children's Hospital, Nijmegen, the Netherlands.

Gijs J Overheul (GJ)

Department of Medical Microbiology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands.

Pascal Miesen (P)

Department of Medical Microbiology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands.

Ronald P van Rij (RP)

Department of Medical Microbiology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands.

Sergio H Triana (SH)

Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany; Broad Institute of MIT and Harvard, Cambridge, MA, USA; Institute for Medical Engineering and Science, MIT, Cambridge, MA, USA; Department of Chemistry, MIT, Cambridge, MA, USA; Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, USA.

Bart Smeets (B)

Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands.

Rebekka K Schneider (RK)

Institute of Cell and Tumor Biology, RWTH Aachen University, Aachen, Germany; Department of Developmental Biology, Erasmus Medisch Centrum, Rotterdam, the Netherlands; Oncode Institute, Erasmus Medisch Centrum, Rotterdam, the Netherlands; Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medisch Centrum, Rotterdam, the Netherlands.

Rafael Kramann (R)

Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, Aachen, Germany; Division of Nephrology and Clinical Immunology, RWTH Aachen University, Aachen, Germany; Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medisch Centrum, Rotterdam, the Netherlands. Electronic address: rkramann@gmx.net.

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Classifications MeSH

questionsmedicales.fr › Maladies de l'appareil respiratoire › Infections de l'appareil respiratoire › Pneumopathie infectieuse › Pneumopathie virale › COVID-19 › Using human iPSC-derived kidney organoids to...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Using human iPSC-derived kidney organoids to...
questionsmedicales.fr › Cellules › Cellules souches › Cellules souches pluripotentes › Cellules souches pluripotentes induites › Using human iPSC-derived kidney organoids to...
questionsmedicales.fr › Appareil urogénital › Voies urinaires › Rein › Using human iPSC-derived kidney organoids to...
questionsmedicales.fr › Tissus › Organoïdes › Using human iPSC-derived kidney organoids to...
questionsmedicales.fr › Virus › Virus à ARN › Virus à ARN à brin positif › Nidovirales › Coronaviridae › Coronavirus › Betacoronavirus › Virus du SRAS › SARS-CoV-2 › Using human iPSC-derived kidney organoids to...