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

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 im...

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Detalles Bibliográficos
Autores principales: Lian, Eric, Pietrobon, Adam, Stanford, William L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10220269/
https://www.ncbi.nlm.nih.gov/pubmed/37220001
http://dx.doi.org/10.1016/j.xpro.2023.102314
Descripción
Sumario: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.(1)