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BRAIN Publication-derived

Human Fetal Brain Organoid (FeBO) Establishment and Long-term Culture

Source Hendriks et al., 2024 · Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands; Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), Utrecht, the Netherlands · 10.1016/j.cell.2023.12.012

👤 Delilah Hendriks, Anna Pagliaro, Francesco Andreatta, Wei Wu, Hans Clevers, Benedetta Artegiani ⏱ 30 days 📋 3 phases 🧫 Human fetal neural stem cells and progenitors (GW12-15)

Abstract

This protocol establishes long-term expanding three-dimensional organoids (FeBOs) from healthy human fetal brain tissue fragments (gestational weeks 12-15). FeBOs self-organize with tissue-like cellular heterogeneity including neural stem cells, radial glia, and neurons, and can be repeatedly passaged while maintaining regional identity and ECM niche properties for CNS development and disease modeling studies.

Cell source
Human fetal neural stem cells and progenitors (GW12-15)
Application
Disease modeling, Developmental study, Drug screening

Protocol overview

16 steps across 3 phases

Tissue Isolation and FeBO Establishment Days 0-8
  1. 1 Obtain human fetal brain tissue
  2. 2 Dissect brain tissue into small fragments
  3. 3 Place tissue fragments in serum-free, extracellular matrix-free expansion medium
  4. 4 Culture on orbital shaker
  5. 5 Monitor FeBO formation
FeBO Passaging and Maintenance Days 8-30+ (Continuous)
  1. 1 Determine optimal passage timing
  2. 2 Harvest organoids
  3. 3 Cut whole organoid into pieces
  4. 4 Transfer organoid pieces to fresh expansion medium
  5. 5 Culture on orbital shaker
  6. 6 Monitor organoid reformation
  7. 7 Establish FeBO lines
Optional: Maturation Phase Days 1-10 (from any passage)
  1. 1 Prepare maturation medium
  2. 2 Switch to maturation medium
  3. 3 Culture in maturation medium
  4. 4 Assess maturation markers

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Hendriks et al., 2024. 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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