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

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 organ...

Descripción completa

Detalles Bibliográficos
Autores principales: Reimer, Katharina C., Jansen, Jitske, Overheul, Gijs J., Miesen, Pascal, van Rij, Ronald P., Triana, Sergio H., Smeets, Bart, Schneider, Rebekka K., Kramann, Rafael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9293950/
https://www.ncbi.nlm.nih.gov/pubmed/35983169
http://dx.doi.org/10.1016/j.xpro.2022.101612
_version_ 1784749749823340544
author Reimer, Katharina C.
Jansen, Jitske
Overheul, Gijs J.
Miesen, Pascal
van Rij, Ronald P.
Triana, Sergio H.
Smeets, Bart
Schneider, Rebekka K.
Kramann, Rafael
author_facet Reimer, Katharina C.
Jansen, Jitske
Overheul, Gijs J.
Miesen, Pascal
van Rij, Ronald P.
Triana, Sergio H.
Smeets, Bart
Schneider, Rebekka K.
Kramann, Rafael
author_sort Reimer, Katharina C.
collection PubMed
description 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).
format Online
Article
Text
id pubmed-9293950
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-92939502022-07-19 Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level Reimer, Katharina C. Jansen, Jitske Overheul, Gijs J. Miesen, Pascal van Rij, Ronald P. Triana, Sergio H. Smeets, Bart Schneider, Rebekka K. Kramann, Rafael STAR Protoc Protocol 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). Elsevier 2022-07-19 /pmc/articles/PMC9293950/ /pubmed/35983169 http://dx.doi.org/10.1016/j.xpro.2022.101612 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Reimer, Katharina C.
Jansen, Jitske
Overheul, Gijs J.
Miesen, Pascal
van Rij, Ronald P.
Triana, Sergio H.
Smeets, Bart
Schneider, Rebekka K.
Kramann, Rafael
Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level
title Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level
title_full Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level
title_fullStr Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level
title_full_unstemmed Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level
title_short Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level
title_sort using human ipsc-derived kidney organoids to decipher sars-cov-2 pathology on single cell level
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9293950/
https://www.ncbi.nlm.nih.gov/pubmed/35983169
http://dx.doi.org/10.1016/j.xpro.2022.101612
work_keys_str_mv AT reimerkatharinac usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT jansenjitske usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT overheulgijsj usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT miesenpascal usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT vanrijronaldp usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT trianasergioh usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT smeetsbart usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT schneiderrebekkak usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel
AT kramannrafael usinghumanipscderivedkidneyorganoidstodeciphersarscov2pathologyonsinglecelllevel

Ejemplares similares