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BRAIN Publication-derived

Forebrain Organoid Differentiation for Modelling Autism Spectrum Disorder

Source Jourdon et al., 2023 · Yale University School of Medicine, Child Study Center · 10.1038/s41593-023-01399-0

👤 Alexandre Jourdon, Feinan Wu, Jessica Mariani, Davide Capauto, Scott Norton, Livia Tomasini, Anahita Amiri, Milovan Suvakov, Jeremy D. Schreiner, Yeongjun Jang, Arijit Panda, Cindy Khanh Nguyen, Elise M. Cummings, Gloria Han, Kelly Powell, Anna Szekely, James C. McPartland, Kevin Pelphrey, Katarzyna Chawarska, Pamela Ventola, Alexej Abyzov, Flora M. Vaccarino ⏱ 100 days 📋 4 phases 🧫 Human iPSC

Abstract

This protocol generates forebrain organoids from human induced pluripotent stem cells (iPSCs) derived from autism spectrum disorder (ASD) patients and unaffected controls. The method produces early-stage forebrain tissues containing diverse neuroectodermal cell lineages—radial glia, intermediate progenitors, and excitatory/inhibitory neurons—enabling transcriptomic and cellular analysis of altered neurodevelopmental trajectories in idiopathic ASD.

Cell source
Human iPSC
Application
Disease modeling

Protocol overview

13 steps across 4 phases

iPSC Preparation and Seeding Day -1 to Day 0
  1. 1 iPSC colony dissociation
  2. 2 Cell counting and seeding in suspension
Neural Induction (Days 1–7) Day 1 to Day 7
  1. 1 Initiate neural induction on Day 1
  2. 2 Switch to KSR medium on Day 2
  3. 3 Transition to NIM on Day 5
  4. 4 Increase NIM concentration on Day 7
Proliferation Phase with Growth Factors (Days 9–16) Day 9 to Day 16
  1. 1 Switch to 100% NIM with FGF and EGF
  2. 2 Maintain FGF/EGF supplementation through Day 16
Terminal Differentiation Phase (Days 17–100) Day 17 (TD0) to Day 100+ (TD30–TD60)
  1. 1 Initiate terminal differentiation at Day 17 (TD0)
  2. 2 Perform half-media changes twice weekly (TD0 to TD30)
  3. 3 Transfer organoids at TD10 to larger culture vessel
  4. 4 Remove growth factors after TD30
  5. 5 Maintain culture through TD60 and beyond

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Jourdon et al., 2023. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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