An efficient method to generate kidney organoids at the air-liquid interface

Ashwani Kumar Gupta; David Z Ivancic; Bilal A Naved; Jason A Wertheim; Leif Oxburgh

doi:10.14440/jbm.2021.357

An efficient method to generate kidney organoids at the air-liquid interface

J Biol Methods. 2021 Jun 30;8(2):e150. doi: 10.14440/jbm.2021.357. eCollection 2021.

Authors

Ashwani Kumar Gupta^{1

2}, David Z Ivancic¹, Bilal A Naved^{1

3}, Jason A Wertheim^{1

2

3

4

5

6}, Leif Oxburgh⁷

Affiliations

¹ Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
² Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
³ Department of Biomedical Engineering, McCormick School of Engineering, Northwestern University, Evanston, IL 60208, USA.
⁴ Simpson Querrey Institute, Northwestern University, Chicago, IL 60611, USA.
⁵ Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208, USA.
⁶ Department of Surgery, Jesse Brown VA Medical Center, Chicago, IL 60612, USA.
⁷ The Rogosin Institute, New York, NY 10021, USA.

Abstract

The prevalence of kidney dysfunction continues to increase worldwide, driving the need to develop transplantable renal tissues. The kidney develops from four major renal progenitor populations: nephron epithelial, ureteric epithelial, interstitial and endothelial progenitors. Methods have been developed to generate kidney organoids but few or dispersed tubular clusters within the organoids hamper its use in regenerative applications. Here, we describe a detailed protocol of asynchronous mixing of kidney progenitors using organotypic culture conditions to generate kidney organoids tightly packed with tubular clusters and major renal structures including endothelial network and functional proximal tubules. This protocol provides guidance in the culture of human embryonic stem cells from a National Institute of Health-approved line and their directed differentiation into kidney organoids. Our 18-day protocol provides a rapid method to generate kidney organoids that facilitate the study of different nephrological events including in vitro tissue development, disease modeling and chemical screening. However, further studies are required to optimize the protocol to generate additional renal-specific cell types, interconnected nephron segments and physiologically functional renal tissues.

Keywords: air-liquid interface; kidney organoids; kidney progenitors; organotypic culture; proximal tubule.

Abstract

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