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

ABSTRACT: 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.

SUBMITTER: Lian E

PROVIDER: S-EPMC10220269 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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

Lian Eric E Pietrobon Adam A Stanford William L WL

STAR protocols 20230522 2

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 ...[more]

PMID: 37220001

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