Cerebral Palsy with Intellectual Disability Brain Organoids
Source Sun et al., 2024 · Guangzhou Women and Children's Medical Center, Guangzhou Medical University · 10.1016/j.isci.2024.109748
Abstract
This protocol establishes a brain organoid differentiation model to investigate how TYW1 loss-of-function impairs neuronal differentiation. TYW1-knockout and wild-type hESCs are differentiated into brain organoids and assessed at 66 days in vitro for neuronal markers (MAP2, β-Tubulin III) and HERVK expression, revealing impaired neural differentiation and elevated endogenous retrovirus expression in knockout cells.
Protocol overview
16 steps across 4 phases
- a Dissociate hESC colonies into single cells
- b Plate cells for EB formation
- c Neural induction with dual SMAD inhibition
- d Media changes during neural induction phase
- a Switch to neural progenitor expansion medium
- b Maintain neural progenitor expansion with daily media changes
- c Continue expansion with every-other-day media changes
- a Switch to neuronal differentiation factors
- b Culture organoids to terminal maturation phase
- c Remove growth factors for terminal maturation
- a Collect organoids
- b Fix organoids for immunostaining
- c Dehydrate and embed in OCT
- d Section organoids for immunostaining
- e Immunostain organoid sections for neuronal markers
- f Quantify neuronal differentiation
Full SOP
Full SOPs need an account and a plan
Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.
Create an accountAlready registered? Log in
Attribution
This SOP was written by Organthis based on the published method in Sun et al., 2024. Once the originating laboratory has confirmed it, the SOP is marked Verified.
Something wrong with this entry? Report an issue with this protocol