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

Autor:	Gupta AK; Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.; Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA., Ivancic DZ; Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA., Naved BA; Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.; Department of Biomedical Engineering, McCormick School of Engineering, Northwestern University, Evanston, IL 60208, USA., Wertheim JA; Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.; Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.; Department of Biomedical Engineering, McCormick School of Engineering, Northwestern University, Evanston, IL 60208, USA.; Simpson Querrey Institute, Northwestern University, Chicago, IL 60611, USA.; Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208, USA.; Department of Surgery, Jesse Brown VA Medical Center, Chicago, IL 60612, USA., Oxburgh L; The Rogosin Institute, New York, NY 10021, USA.
Jazyk:	angličtina
Zdroj:	Journal of biological methods [J Biol Methods] 2021 Jun 30; Vol. 8 (2), pp. e150. Date of Electronic Publication: 2021 Jun 30 (Print Publication: 2021).
DOI:	10.14440/jbm.2021.357
Abstrakt:	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. (© 2013-2021 The Journal of Biological Methods, All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu