Self-Organizing Human Brain Tissue
Source Lancaster et al. · IMBA — Institute of Molecular Biotechnology, Vienna, Austria · 10.1038/nature12517
Abstract
Landmark publication establishing the first protocol for generating cerebral organoids from human iPSCs through self-organized differentiation in Matrigel droplets and spinning bioreactors. Lancaster et al. demonstrated that these 3D neural tissues recapitulate features of early human cortical development and successfully modeled primary microcephaly using patient-derived iPSCs with CDK5RAP2 mutations.
Protocol overview
17 steps across 5 phases
- 1 Dissociate and prepare pluripotent stem cells
- 2 Plate cells in ultra-low-binding wells
- 3 Culture embryoid bodies
- 1 Transfer to neural induction media
- 2 Culture neuroectodermal tissues
- 1 Prepare Matrigel droplets
- 2 Allow Matrigel to gel and transfer to culture
- 3 Culture in stationary differentiation media
- 4 Maintain in stationary culture for 4 days
- 1 Transfer to spinning bioreactor
- 2 Establish bioreactor culture conditions
- 3 Feed and monitor throughout culture
- 4 Assess brain region development
- 1 Prepare electroporation apparatus
- 2 Inject plasmid DNA into organoid
- 3 Perform electroporation
- 4 Return organoid to culture
Full SOP
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Attribution
This SOP was authored by Organthis based on the published method in Lancaster et al.. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.
This wording is awaiting legal review.
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