Transient Hypoxic Injury Model in Human Cerebral Organoids
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
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.
Protocol overview
12 steps across 5 phases
- 1 Plate H9 hESCs in ultra-low attachment 96-well plates
- 2 Initiate neural induction
- 1 Embed organoids in Matrigel droplets
- 2 Culture in 6 cm Petri dishes for stationary phase
- 1 Transfer organoids to orbital shaker
- 2 Maintain rotating culture until hypoxia induction
- 1 Transfer organoids to hypoxic chamber
- 2 Maintain hypoxic conditions for 24 hours
- 3 Return organoids to normoxic conditions
- 1 Continue rotating culture during recovery phase
- 2 Perform cell labeling studies at defined timepoints
- 3 Collect organoids for analysis
Full SOP
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Attribution
This SOP was authored by Organthis based on the published method in Daviaud et al., 2019. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.
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