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BRAIN Publication-derived

Brain Assembloids for Disease Modelling with Interneurons

Full SOP with Academic or a plan

Source Birey et al. · Stanford University School of Medicine, Stanford, CA, USA · 10.1038/nature22330

Authors: Birey F, Andersen J, Makinson CD, Islam S, Wei W, Huber N, Fan HC, Mber KRC, Bhatt SM, Bhatt AH, Pasca AM, Paşca SP Duration: 90 days 4 phases Cell source: Human iPSC

Abstract

Birey et al. published the landmark assembloid study fusing cortical (hCS) and subpallial (hSS) spheroids to demonstrate saltatory interneuron migration from ventral to dorsal forebrain organoids, recapitulating a key developmental process. Using Timothy Syndrome patient iPSCs, they showed that CACNA1C mutations impair this migration, establishing assembloids as a paradigm for modeling circuit assembly disorders.

Cell source
Human iPSC
Application
Interneuron Migration Studies

Protocol overview

11 steps across 4 phases

Phase 1 · hPS cell culture and initiation of neural spheroid formation Days 0-6
  1. a Culture hPS cells on feeder layer
  2. b Prepare hPS cell medium
  3. c Lift and transfer hPS cell colonies
  4. d Initiate neural induction with SMAD inhibitors
  5. e Transition to neural medium at day 6
  6. f Add pathway modulators for hSS generation (hSS only)
Phase 2 · Growth factor supplementation and neuronal maturation Days 25-42
  1. a Add neurotrophic factors
Phase 3 · Long-term culture and maintenance Days 43 onwards
  1. a Maintain in unsupplemented neural medium
Phase 4 · Viral labeling and spheroid assembly Days 60-90
  1. a Perform viral infection of spheroids (8-10 days before assembly)
  2. b Assemble forebrain spheroids
  3. c Transfer fused spheroids to culture plates

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Attribution

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

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