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Abstract

Developmental Biology

Generating Kidney Organoids in Suspension from Induced Pluripotent Stem Cells

Published: September 1st, 2023

DOI:

10.3791/65698

1Department of Nephrology, The Children's Hospital, Zhejiang University School of Medicine, 2Zhejiang University School of Medicine, 3Hubei Normal University, 4National Clinical Research Center of Kidney Diseases, Jinling Hospital, Nanjing University School of Medicine, 5Clinical Laboratory, The Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health

* These authors contributed equally

Abstract

Kidney organoids can be generated from induced pluripotent stem cells (iPSCs) through various approaches. These organoids hold great promise for disease modeling, drug screening, and potential therapeutic applications. This article presents a step-by-step procedure to create kidney organoids from iPSCs, starting from the posterior primitive streak (PS) to the intermediate mesoderm (IM). The approach relies on the APEL 2 medium, which is a defined, animal component-free medium. It is supplemented with a high concentration of WNT agonist (CHIR99021) for a duration of 4 days, followed by fibroblast growth factor 9 (FGF9)/heparin and a low concentration of CHIR99021 for an additional 3 days. During this process, emphasis is given to selecting the optimal cell density and CHIR99021 concentration at the start of iPSCs, as these factors are critical for successful kidney organoid generation. An important aspect of this protocol is the suspension culture in a low adherent plate, allowing the IM to gradually develop into nephron structures, encompassing glomerular, proximal tubular, and distal tubular structures, all presented in a visually comprehensible format. Overall, this detailed protocol offers an efficient and specific technique to produce kidney organoids from diverse iPSCs, ensuring successful and consistent results.

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Keywords Induced Pluripotent Stem Cells

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