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BRAIN Publication-derived

Miller-Dieker Syndrome Cortical Organoids for Disease Modelling

Full SOP with Academic or a plan

Source Iefremova et al., 2017 · Institute of Reconstructive Neurobiology, University of Bonn · 10.1016/j.celrep.2017.03.047

Authors: Vira Iefremova, George Manikakis, Olivia Krefft, Ammar Jabali, Kevin Weynans, Ruven Wilkens, Fabio Marsoner, Björn Brändl, Franz-Josef Müller, Philipp Koch, Julia Ladewig Duration: 35 days 3 phases Cell source: Patient-Derived iPSC (Miller-Dieker Syndrome)

Abstract

This protocol generates Miller-Dieker-syndrome-specific iPSC-derived forebrain-type organoids that model cortical development and disease-related defects. The organoids exhibit reduced expansion rates, premature neurogenesis, and disrupted cortical niche architecture due to alterations in the LIS1/NDEL1/14.3.3ε complex. The model reveals non-cell-autonomous defects in Wnt/β-catenin signaling that can be rescued by GSK3β inhibition.

Cell source
Patient-Derived iPSC (Miller-Dieker Syndrome)
Application
Disease modeling

Protocol overview

10 steps across 3 phases

Phase 1 · iPSC Dissociation and Embryoid Body Formation Day 1-5
  1. a Dissociate iPSCs into single cells
  2. b Exchange medium every other day
  3. c Transfer embryoid bodies to low-adhesion plates
Phase 2 · Neural Induction and Small Molecule Treatment Day 5-12
  1. a Maintain neural induction for 5 days with small molecules
  2. b Remove small molecules from medium
  3. c Embed organoids in Geltrex matrix
Phase 3 · Organoid Culture and Maturation Day 12-35+
  1. a Culture organoids under continuous agitation
  2. b Change medium every 3-4 days
  3. c Optional: Add GSK3β inhibitor for β-catenin/Wnt activation
  4. d Harvest organoids for analysis

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Attribution

This SOP was written by Organthis based on the published method in Iefremova et al., 2017. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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