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BRAIN Publication-derived

Alzheimer's Disease Cerebral Organoids for Drug Screening (2022)

Full SOP with Academic or a plan

Source Qiao et al., 2022 · Wuhan University TaiKang Medical School, Department of Biomedical Engineering · 10.3390/ijms23115981

Authors: Haowen Qiao, Wen Zhao, Moujian Guo, Lili Zhu, Tao Chen, Jibo Wang, Xiaodong Xu, Zhentao Zhang, Ying Wu, Pu Chen Duration: 65 days 5 phases Cell source: Human ESC (H9)

Abstract

This protocol generates human cerebral organoids from H9 embryonic stem cells to model HSV-1-induced Alzheimer's disease-associated neuropathology, including amyloid-beta deposition, neuroinflammation, and neuron loss. The organoids are used to evaluate therapeutic potential of antiviral drugs (Ribavirin and Valacyclovir) in rescuing disease phenotypes, providing a human-relevant in vitro infection model for mechanistic study and drug discovery.

Cell source
Human ESC (H9)
Application
Disease modeling; drug screening

Protocol overview

19 steps across 5 phases

Phase 1 · Embryoid Body Formation and Initial Differentiation Days 0-12
  1. a Prepare hESC suspension
  2. b Form embryoid bodies (EBs)
  3. c Transfer EBs to ultra-low-attachment plates
  4. d Feed EBs and transition to induction
Phase 2 · Matrigel Embedding and Neural Expansion Days 12-28
  1. a Embed EBs in Matrigel droplets
  2. b Culture in neural expansion medium
  3. c Transfer to orbital shaker
Phase 3 · HSV-1 Infection Days 14, 42, or 65 (+ 3 days infection)
  1. a Prepare HSV-1 inoculum
  2. b Infect organoids with HSV-1
  3. c Wash organoids after infection
Phase 4 · Antiviral Drug Treatment (Optional Phenotypic Rescue) Days 14, 42, or 65 (+ 3 days treatment)
  1. a Prepare antiviral drug treatments
  2. b Co-incubate organoids with HSV-1 and antiviral drugs
  3. c Wash organoids after drug treatment
Phase 5 · Sample Preparation and Analysis Days 14+3, 42+3, 65+3 (post-infection/treatment)
  1. a Prepare organoids for immunofluorescence
  2. b Perform immunofluorescence staining
  3. c Image organoid sections
  4. d Perform whole-mount 3D immunostaining (optional for select markers)
  5. e Extract RNA for RT-PCR analysis
  6. f Perform real-time RT-PCR

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Attribution

This SOP was written by Organthis based on the published method in Qiao et al., 2022. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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