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Cell Stem Cell. 2018 Dec 6;23(6):869-881.e8. doi: 10.1016/j.stem.2018.10.010. Epub 2018 Nov 15.

Comparative Analysis and Refinement of Human PSC-Derived Kidney Organoid Differentiation with Single-Cell Transcriptomics.

Author information

1
Division of Nephrology, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA.
2
Department of Developmental Biology, Washington University School of Medicine, St. Louis, MO, USA; Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA.
3
Division of Nephrology, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA; Department of Developmental Biology, Washington University School of Medicine, St. Louis, MO, USA. Electronic address: humphreysbd@wustl.edu.

Abstract

Kidney organoids derived from human pluripotent stem cells have great utility for investigating organogenesis and disease mechanisms and, potentially, as a replacement tissue source, but how closely organoids derived from current protocols replicate adult human kidney is undefined. We compared two directed differentiation protocols by single-cell transcriptomics of 83,130 cells from 65 organoids with single-cell transcriptomes of fetal and adult kidney cells. Both protocols generate a diverse range of kidney cells with differing ratios, but organoid-derived cell types are immature, and 10%-20% of cells are non-renal. Reconstructing lineage relationships by pseudotemporal ordering identified ligands, receptors, and transcription factor networks associated with fate decisions. Brain-derived neurotrophic factor (BDNF) and its cognate receptor NTRK2 were expressed in the neuronal lineage during organoid differentiation. Inhibiting this pathway improved organoid formation by reducing neurons by 90% without affecting kidney differentiation, highlighting the power of single-cell technologies to characterize and improve organoid differentiation.

KEYWORDS:

BDNF signaling; Dropseq; human kidney; induced pluripotent stem cells; kidney organoid; off-target cells; single cell RNA-seq; stem cell differentiation

PMID:
30449713
PMCID:
PMC6324730
[Available on 2019-12-06]
DOI:
10.1016/j.stem.2018.10.010

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