title: Generation of kidney organoids from human pluripotent stem cells.
abstract: 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.
authors: Murdoch Childrens Research Institute, Parkville, Melbourne, Victoria, Australia.|Institute for Molecular Bioscience, The University of Queensland, St Lucia, Queensland, Australia.
authors: Minoru
authors: 0000-0002-0458-7414
authors: M
authors: Takasato
authors: Murdoch Childrens Research Institute, Parkville, Melbourne, Victoria, Australia.|Institute for Molecular Bioscience, The University of Queensland, St Lucia, Queensland, Australia.
authors: Pei X
authors: PX
authors: Er
authors: Institute for Molecular Bioscience, The University of Queensland, St Lucia, Queensland, Australia.
authors: Han S
authors: HS
authors: Chiu
authors: Murdoch Childrens Research Institute, Parkville, Melbourne, Victoria, Australia.|Institute for Molecular Bioscience, The University of Queensland, St Lucia, Queensland, Australia.|Department of Paediatrics, The University of Melbourne, Parkville, Victoria, Australia.
authors: Melissa H
authors: MH
authors: Little
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007668:Kidney / Q000166:cytology*; D009940:Organoids / Q000166:cytology*