title: Making a Kidney Organoid Using the Directed Differentiation of Human Pluripotent Stem Cells.
abstract: An organoid can be defined as a three-dimensional organ-like structure formed from organ-specific progenitor cells. Organ progenitor cells were empirically found to self-organize three-dimensional tissues when they were aggregated and cultivated in vitro. While this nature power of progenitor cells has an amazing potential to recreate artificial organs in vitro, there had been difficulty to apply this technology to human organs due to the inaccessibility to human progenitor cells until human-induced pluripotent stem cell (hiPSC) was invented by Takahashi and Yamanaka in 2007. As embryonic stem cells do, hiPSCs also have pluripotency to give rise to any organs/tissues cell types, including the kidney, via directed differentiation. Here, we provide a detailed protocol for generating kidney organoids using human pluripotent stem cells. The protocol differentiates human pluripotent stem cells into the posterior primitive streak. This is followed by the simultaneous induction of posterior and anterior intermediate mesoderm that are subsequently aggregated and undergo self-organization into the kidney organoid. Such kidney organoids are comprised of all anticipated kidney cell types including nephrons segmented into the glomerulus, proximal tubule, loop of Henle, and distal tubule as well as the collecting duct, endothelial network, and renal interstitium.
authors.affiliation: Murdoch Children's Research Institute, Flemington Road, Parkville, VIC, 3052, Australia. minoru.takasato@riken.jp.|RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuou-ku, Kobe, Hyogo, 650-0047, Japan. minoru.takasato@riken.jp.
authors.forename: Minoru
authors.initials: M
authors.lastname: Takasato
authors.affiliation: Murdoch Children's Research Institute, Flemington Road, Parkville, VIC, 3052, Australia.|Department of Pediatrics, University of Melbourne, Parkville, VIC, 3010, Australia.
authors.forename: Melissa H
authors.initials: MH
authors.lastname: Little
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007668:Kidney / Q000166:cytology*; D008648:Mesoderm; D051379:Mice; D009940:Organoids / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*
keywords: Directed differentiation; Early embryogenesis; Human pluripotent stem cells; Intermediate mesoderm; Kidney development; Kidney organoid; Metanephric mesenchyme; Primitive streak; Ureteric bud