Generation of kidney organoids from human pluripotent stem cells
- 18 August 2016
- journal article
- research article
- Published by Springer Nature in Nature Protocols
- Vol. 11 (9), 1681-1692
- https://doi.org/10.1038/nprot.2016.098
Abstract
This protocol describes stepwise differentiation of human pluripotent stem cells into 3D kidney organoids that contain segmented nephrons connected to collecting ducts, which are surrounded by renal interstitial cells and an endothelial network. The human kidney develops from four progenitor populations—nephron progenitors, ureteric epithelial progenitors, renal interstitial progenitors and endothelial progenitors—resulting in the formation of maximally 2 million nephrons. Until recently, the reported methods differentiated human pluripotent stem cells (hPSCs) into either nephron progenitor or ureteric epithelial progenitor cells, consequently forming only nephrons or collecting ducts, respectively. Here we detail a protocol that simultaneously induces all four progenitors to generate kidney organoids within which segmented nephrons are connected to collecting ducts and surrounded by renal interstitial cells and an endothelial network. As evidence of functional maturity, proximal tubules within organoids display megalin-mediated and cubilin-mediated endocytosis, and they respond to a nephrotoxicant to undergo apoptosis. This protocol consists of 7 d of monolayer culture for intermediate mesoderm induction, followed by 18 d of 3D culture to facilitate self-organizing renogenic events leading to organoid formation. Personnel experienced in culturing hPSCs are required to conduct this protocol.Keywords
This publication has 51 references indexed in Scilit:
- Self-Formation of Optic Cups and Storable Stratified Neural Retina from Human ESCsCell Stem Cell, 2012
- Directed differentiation of human pluripotent stem cells into intestinal tissue in vitroNature, 2010
- Genetic kidney diseasesThe Lancet, 2010
- Differentiation of Neuroepithelia from Human Embryonic Stem CellsPublished by Springer Nature ,2009
- Osr1 expression demarcates a multi-potent population of intermediate mesoderm that undergoes progressive restriction to an Osr1-dependent nephron progenitor compartment within the mammalian kidneyDevelopmental Biology, 2008
- Six2 Defines and Regulates a Multipotent Self-Renewing Nephron Progenitor Population throughout Mammalian Kidney DevelopmentCell Stem Cell, 2008
- The migration of paraxial and lateral plate mesoderm cells emerging from the late primitive streak is controlled by different Wnt signalsBMC Developmental Biology, 2008
- Hoxd11 specifies a program of metanephric kidney development within the intermediate mesoderm of the mouse embryoDevelopmental Biology, 2008
- A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cellsProceedings of the National Academy of Sciences, 2007
- Wnt and TGF-β signaling are required for the induction of an in vitro model of primitive streak formation using embryonic stem cellsProceedings of the National Academy of Sciences, 2006