An Organoid-Based Model of Cortical Development Identifies Non-Cell-Autonomous Defects in Wnt Signaling Contributing to Miller-Dieker Syndrome
Source Iefremova et al., 2017 · Institute of Reconstructive Neurobiology, University of Bonn · 10.1016/j.celrep.2017.03.047
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.
Protocol overview
10 steps across 3 phases
- 1 Dissociate iPSCs into single cells
- 2 Exchange medium every other day
- 3 Transfer embryoid bodies to low-adhesion plates
- 4 Maintain neural induction for 5 days with small molecules
- 5 Remove small molecules from medium
- 6 Embed organoids in Geltrex matrix
- 7 Culture organoids under continuous agitation
- 8 Change medium every 3–4 days
- 9 Optional: Add GSK3β inhibitor for β-catenin/Wnt activation
- 10 Harvest organoids for analysis
Full SOP
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Attribution
This SOP was authored by Organthis based on the published method in Iefremova et al., 2017. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.
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