A CRISP(e)R view on kidney organoids allows generation of an induced pluripotent stem cell-derived kidney model for drug discovery

Kidney Int. 2018 Dec;94(6):1099-1110. doi: 10.1016/j.kint.2018.05.003. Epub 2018 Jul 31.


Development of physiologically relevant cellular models with strong translatability to human pathophysiology is critical for identification and validation of novel therapeutic targets. Herein we describe a detailed protocol for generation of an advanced 3-dimensional kidney cellular model using induced pluripotent stem cells, where differentiation and maturation of kidney progenitors and podocytes can be monitored in live cells due to CRISPR/Cas9-mediated fluorescent tagging of kidney lineage markers (SIX2 and NPHS1). Utilizing these cell lines, we have refined the previously published procedures to generate a new, higher throughput protocol suitable for drug discovery. Using paraffin-embedded sectioning and whole-mount immunostaining, we demonstrated that organoids grown in suspension culture express key markers of kidney biology (WT1, ECAD, LTL, nephrin) and vasculature (CD31) within renal cortical structures with microvilli, tight junctions and podocyte foot processes visualized by electron microscopy. Additionally, the organoids resemble the adult kidney transcriptomics profile, thereby strengthening the translatability of our in vitro model. Thus, development of human nephron-like structures in vitro fills a major gap in our ability to assess the effect of potential treatment on key kidney structures, opening up a wide range of possibilities to improve clinical translation.

Keywords: glomerulus; kidney development; podocyte; proximal tubule; stem cell.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Biomarkers / metabolism
  • CRISPR-Cas Systems*
  • Cell Differentiation
  • Cell Line
  • Cell Lineage
  • Drug Discovery / methods*
  • Gene Editing / methods*
  • Gene Expression Regulation
  • Genotype
  • High-Throughput Screening Assays
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / physiology*
  • Induced Pluripotent Stem Cells / ultrastructure
  • Kidney / drug effects
  • Kidney / metabolism
  • Kidney / physiology*
  • Kidney / ultrastructure
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Organoids / drug effects
  • Organoids / metabolism
  • Organoids / physiology*
  • Organoids / ultrastructure
  • Phenotype
  • Podocytes / drug effects
  • Podocytes / metabolism
  • Podocytes / physiology*
  • Podocytes / ultrastructure
  • Time Factors
  • Transcriptome


  • Biomarkers
  • Homeodomain Proteins
  • Membrane Proteins
  • Nerve Tissue Proteins
  • SIX2 protein, human
  • nephrin