Skip to content
← Back to browse
BRAIN Publication-derived

Fused Cerebral Organoids Model Interactions Between Brain Regions

Source Bagley et al., 2017 · Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna, Austria · 10.1038/nmeth.4304

👤 Joshua A Bagley, Daniel Reumann, Shan Bian, Julie Lévi-Strauss, Juergen A Knoblich ⏱ 80 days 📋 7 phases 🧫 Human iPSC

Abstract

This protocol describes a method for generating fused dorsal and ventral forebrain cerebral organoids from human pluripotent stem cells to model long-distance GABAergic interneuron migration within a single tissue construct. The fused organoid system recapitulates aspects of human brain development and enables functional studies of neuronal migration relevant to neurodevelopmental disorders.

Cell source
Human iPSC
Application
Disease modeling

Protocol overview

22 steps across 7 phases

hPSC Maintenance and Preparation Days -14 to 0
  1. 1 Culture feeder-dependent hiPSCs
  2. 2 Maintain feeder-free H9 hESCs
  3. 3 Prepare hPSCs for embryoid body formation
Embryoid Body (EB) Formation and Neural Induction Days 0–5
  1. 1 Generate embryoid bodies in suspension
  2. 2 Initiate neural induction with hES medium + bFGF and RI media
Drug Patterning Treatment (Days 5–11) Days 5–11
  1. 1 Apply dorsal patterning (untreated control)
  2. 2 Apply ventral patterning (WNT inhibition + SHH enhancement)
  3. 3 Apply dorsal[CycA] patterning (enhanced dorsal identity)
Organoid Embedding and Growth (Days 12–30) Days 12–30
  1. 1 Embed EBs in Matrigel droplets
  2. 2 Maintain organoids on orbital shaker
  3. 3 Supplement medium with Matrigel after day 40
Organoid Maturation and Analysis (Days 30–80+) Days 30–80+
  1. 1 Perform qPCR analysis at day 30–40
  2. 2 Harvest organoids for cryosectioning and immunostaining
  3. 3 Perform immunofluorescence on tissue sections
  4. 4 Perform whole-mount and tile-scan imaging
Time-Lapse Imaging of Cell Migration (Days 45–50) Days 45–50
  1. 1 Prepare organoid fusion slice cultures
  2. 2 Perform time-lapse confocal imaging
  3. 3 Analyze cell migration dynamics
Pharmacological Perturbation (3-Week Slice Culture) Days 30–50 (as initiated)
  1. 1 Prepare organoid fusion slice cultures for drug treatment
  2. 2 Apply CXCR4 inhibitor treatment
  3. 3 Fix and immunostain treated slice cultures
  4. 4 Quantify migrated cell density

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

This SOP was authored by Organthis based on the published method in Bagley et al., 2017. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence