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BRAIN Publication-derived

Brain Organoids for Disease Modelling

Full SOP with Academic or a plan

Source Lancaster et al. · IMBA — Institute of Molecular Biotechnology, Vienna, Austria · 10.1038/nature12517

Authors: Lancaster MA, Renner M, Martin CA, Wenzel D, Bicknell LS, Hurles ME, Homfray T, Penninger JM, Jackson AP, Knoblich JA Duration: 30 days 5 phases Cell source: Human iPSC / Human ESC

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.

Cell source
Human iPSC / Human ESC
Application
Neurodevelopmental Disease Modeling

Protocol overview

17 steps across 5 phases

Phase 1 · Embryoid Body Formation Day 0-6
  1. a Dissociate and prepare pluripotent stem cells
  2. b Plate cells in ultra-low-binding wells
  3. c Culture embryoid bodies
Phase 2 · Neural Induction Day 6-11
  1. a Transfer to neural induction media
  2. b Culture neuroectodermal tissues
Phase 3 · Matrigel Embedding and Bioreactor Transfer Day 11-15
  1. a Prepare Matrigel droplets
  2. b Allow Matrigel to gel and transfer to culture
  3. c Culture in stationary differentiation media
  4. d Maintain in stationary culture for 4 days
Phase 4 · Bioreactor Culture and Brain Region Development Day 15-30 days and beyond (up to 10 months)
  1. a Transfer to spinning bioreactor
  2. b Establish bioreactor culture conditions
  3. c Feed and monitor throughout culture
  4. d Assess brain region development
Phase 5 · Optional: Electroporation for Molecular Manipulation Day 12 onwards
  1. a Prepare electroporation apparatus
  2. b Inject plasmid DNA into organoid
  3. c Perform electroporation
  4. d Return organoid to culture

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Attribution

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

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