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BRAIN Publication-derived

Prenatal Hypoxic Injury Brain Organoids

Full SOP with Academic or a plan

Source Daviaud et al., 2019 · Nash Family Department of Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai · 10.3389/fncel.2019.00336

Authors: Nicolas Daviaud, Clément Chevalier, Roland H. Friedel, Hongyan Zou Duration: 42 days 5 phases Cell source: Human ESC (H9)

Abstract

This protocol generates dorsal forebrain-specified cerebral organoids from human H9 embryonic stem cells and models transient prenatal hypoxic injury via 24-hour exposure to 3% O₂ at day 28 of differentiation. The model allows investigation of neuroprogenitor subtype vulnerability, including differential responses of outer radial glia, intermediate progenitors, and neural stem cells to hypoxic stress during early human corticogenesis.

Cell source
Human ESC (H9)
Application
Disease modeling - Prenatal hypoxic injury and neurodevelopmental disorders

Protocol overview

12 steps across 5 phases

Phase 1 · Embryoid Body Formation and Neural Induction D0-D14
  1. a Plate H9 hESCs in ultra-low attachment 96-well plates
  2. b Initiate neural induction
Phase 2 · Matrigel Embedding and Initial Culture D14-D18
  1. a Embed organoids in Matrigel droplets
  2. b Culture in 6 cm Petri dishes for stationary phase
Phase 3 · Orbital Shaker Culture and Maturation D18-D28
  1. a Transfer organoids to orbital shaker
  2. b Maintain rotating culture until hypoxia induction
Phase 4 · Transient Hypoxic Injury Induction D28-D29
  1. a Transfer organoids to hypoxic chamber
  2. b Maintain hypoxic conditions for 24 hours
  3. c Return organoids to normoxic conditions
Phase 5 · Post-Hypoxia Recovery and Analysis D29-D42
  1. a Continue rotating culture during recovery phase
  2. b Perform cell labeling studies at defined timepoints
  3. c Collect organoids for analysis

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Attribution

This SOP was written by Organthis based on the published method in Daviaud et al., 2019. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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