Differentiation and single-cell RNA-seq analyses of human pluripotent-stem-cell-derived renal organoids.

CRISPR Cell Biology Cell Culture Cell Differentiation Genomics Molecular Biology Organoids RNA-seq Sequence Analysis 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:
22 May 2023
Historique:
received: 08 02 2023
revised: 24 03 2023
accepted: 25 04 2023
medline: 23 5 2023
pubmed: 23 5 2023
entrez: 23 5 2023
Statut: aheadofprint

Résumé

Here, we present a protocol for the maintenance and differentiation of human pluripotent stem cells into renal organoids. We describe steps for using a series of readily made differentiation media, multiplexed sample single-cell RNA-seq analysis, quality control, and validation of organoids using immunofluorescence. This provides a rapid and reproducible model of human kidney development and renal disease modeling. Finally, we detail genome engineering using CRISPR-Cas9 homology-directed repair for the generation of renal disease models. For complete details on the use and execution of this protocol, please refer to Pietrobon et al.

Identifiants

DOI: 10.1016/j.xpro.2023.102314 PMID: 37220001 PMC: PMC10220269
pubmed: 37220001
pii: S2666-1667(23)00281-2
doi: 10.1016/j.xpro.2023.102314
pmc: PMC10220269
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

102314

Informations de copyright

Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Références

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Auteurs

Eric Lian (E)

The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada; Ottawa Institute of Systems Biology, Ottawa, ON K1H 8L1, Canada. Electronic address: elian@ohri.ca.

Adam Pietrobon (A)

The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada; Ottawa Institute of Systems Biology, Ottawa, ON K1H 8L1, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8L1, Canada.

William L Stanford (WL)

The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada; Ottawa Institute of Systems Biology, Ottawa, ON K1H 8L1, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8L1, Canada; Department of Biochemistry, Microbiology & Immunology, University of Ottawa, Ottawa, ON K1H 8L1, Canada. Electronic address: wstanford@ohri.ca.

Classifications MeSH