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BRAIN Publication-derived

Cerebral Organoids for Modeling of HSV-1-Induced-Amyloid β Associated Neuropathology and Phenotypic Rescue

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

👤 Haowen Qiao, Wen Zhao, Moujian Guo, Lili Zhu, Tao Chen, Jibo Wang, Xiaodong Xu, Zhentao Zhang, Ying Wu, Pu Chen ⏱ 65 days 📋 5 phases 🧫 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

Embryoid Body Formation and Initial Differentiation Days 0-12
  1. 1 Prepare hESC suspension
  2. 2 Form embryoid bodies (EBs)
  3. 3 Transfer EBs to ultra-low-attachment plates
  4. 4 Feed EBs and transition to induction
Matrigel Embedding and Neural Expansion Days 12-28
  1. 5 Embed EBs in Matrigel droplets
  2. 6 Culture in neural expansion medium
  3. 7 Transfer to orbital shaker
HSV-1 Infection Days 14, 42, or 65 (+ 3 days infection)
  1. 8 Prepare HSV-1 inoculum
  2. 9 Infect organoids with HSV-1
  3. 10 Wash organoids after infection
Antiviral Drug Treatment (Optional Phenotypic Rescue) Days 14, 42, or 65 (+ 3 days treatment)
  1. 11 Prepare antiviral drug treatments
  2. 12 Co-incubate organoids with HSV-1 and antiviral drugs
  3. 13 Wash organoids after drug treatment
Sample Preparation and Analysis Days 14+3, 42+3, 65+3 (post-infection/treatment)
  1. 14 Prepare organoids for immunofluorescence
  2. 15 Perform immunofluorescence staining
  3. 16 Image organoid sections
  4. 17 Perform whole-mount 3D immunostaining (optional for select markers)
  5. 18 Extract RNA for RT-PCR analysis
  6. 19 Perform real-time RT-PCR

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Qiao et al., 2022. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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