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

Journal article
Authors C. Borestrom
A. Jonebring
J. Guo
H. Palmgren
L. Cederblad
A. Forslow
A. Svensson
M. Soderberg
A. Reznichenko
Jenny Nyström
J. Patrakka
R. Hicks
M. Maresca
B. Valastro
A. Collen
Published in Kidney International
Volume 94
Issue 6
Pages 1099-1110
ISSN 0085-2538
Publication year 2018
Published at Institute of Neuroscience and Physiology
Pages 1099-1110
Language en
Keywords glomerulus, kidney development, podocyte, proximal tubule, stem cell, rna-seq, epithelial-cells, podocytes, specificity, injury, cas9, differentiation, proteinuria, mechanisms, crosstalk, Urology & Nephrology
Subject categories Internal medicine


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.

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