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KIDNEY Publication-derived

Kidney Organoids for Drug Screening and Toxicity Testing

Full SOP with Academic or a plan

Source Takasato et al., 2015 · Murdoch Childrens Research Institute, The Royal Children's Hospital Melbourne · 10.1038/nature15695

Authors: Minoru Takasato, Pei X. Er, Han S. Chiu, Barbara Maier, Gregory J. Baillie, Charles Ferguson, Robert G. Parton, Ernst J. Wolvetang, Matthias S. Roost, Susana M. Chuva de Sousa Lopes, Melissa H. Little Duration: 27 days 5 phases Cell source: Human iPSC

Abstract

This protocol generates complex kidney organoids from human induced pluripotent stem cells (iPSCs) that contain multiple nephron lineages and supporting tissues. The organoids model human nephrogenesis and develop functional proximal tubules capable of endocytosis and response to nephrotoxic compounds.

Cell source
Human iPSC
Application
Disease modeling, Developmental study, Drug screening (nephrotoxicity)

Protocol overview

14 steps across 5 phases

Phase 1 · iPSC Expansion and Monolayer Culture Preparation -14 to 0
  1. a Maintain undifferentiated human iPS cells on feeder layer
  2. b Passage cells onto Matrigel and establish monolayer
Phase 2 · Monolayer Differentiation to Intermediate Mesoderm 0 to 7
  1. a Initiate Wnt signaling with CHIR99021
  2. b Add FGF9 and heparin to induce differentiation
  3. c Optional: Add retinoic acid or RA receptor antagonist
  4. d Collect and dissociate cells at day 7
Phase 3 · 3D Organoid Culture and Nephrogenesis Induction 7 to 27
  1. a Form cell aggregates (pellets) on Transwell membrane
  2. b Apply CHIR99021 pulse to trigger nephrogenesis
  3. c Culture organoids with FGF9 and heparin
  4. d Culture organoids in basal medium for final maturation
Phase 4 · Alternative: Monolayer Culture for Lineage Optimization 0 to 16
  1. a Culture cells in monolayer with FGF9 and heparin
  2. b Culture monolayer cells in basal medium
Phase 5 · Functional Analysis and Maturation Assessment 17 to 18
  1. a Assess proximal tubule endocytosis capacity
  2. b Test nephrotoxicity response to cisplatin

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Attribution

This SOP was written by Organthis based on the published method in Takasato et al., 2015. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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