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BRAIN Publication-derived

Cortical-Blood Vessel Assembloids for Modeling SARS-CoV-2-Induced Alzheimer's Disease Pathology

Source Kong et al., 2023 · Adult Stem Cell Research Center and Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University · 10.1038/s41420-022-01288-8

👤 Dasom Kong, Ki Hoon Park, Da-Hyun Kim, Nam Gyo Kim, Seung-Eun Lee, Nari Shin, Myung Geun Kook, Young Bong Kim, Kyung-Sun Kang ⏱ 71 days 📋 4 phases 🧫 Human iPSC (KSCBi005-A)

Abstract

This protocol describes the generation of fused cortical-blood vessel organoids (fCBOs) from human iPSC-derived cortical organoids and blood vessel organoids. The fCBOs provide vascularized brain tissue with integrated immune cells (microglia and astrocytes) to model SARS-CoV-2 infection and associated Alzheimer's disease phenotypes including β-amyloid plaques and tau phosphorylation.

Cell source
Human iPSC (KSCBi005-A)
Application
Disease modeling - SARS-CoV-2 neurotropism and Alzheimer's disease pathology in brain organoids

Protocol overview

20 steps across 4 phases

Cortical Organoid (CO) Generation and Culture Days 0-57
  1. 1 iPSC dissociation and embryoid body formation
  2. 2 Medium change for dual-SMAD inhibition
  3. 3 Transition to neural differentiation medium
  4. 4 Trophic factor supplementation and medium switching
  5. 5 Final maturation culture
Blood Vessel Organoid (BVO) Generation and Culture Days 0-15
  1. 1 iPSC aggregation with BMP4 and CHIR99021
  2. 2 VEGF and forskolin treatment for vascular commitment
  3. 3 Collagen embedding and StemPro culture
  4. 4 Blood vessel network extraction
  5. 5 BVN aggregation into self-assembled BVOs
Fusion of Cortical and Blood Vessel Organoids (fCBO Generation) Days 25-71 (co-culture after fusion)
  1. 1 Prepare cortical and blood vessel organoids for fusion
  2. 2 Co-culture and physical fusion
  3. 3 Transfer to spinning culture
  4. 4 Scaling up and long-term spinning culture
  5. 5 Characterization and validation
SARS-CoV-2 Infection and Analysis Day 57 onwards
  1. 1 Prepare fCBOs for viral infection
  2. 2 SARS-CoV-2 inoculation
  3. 3 Post-infection culture
  4. 4 Mock control preparation
  5. 5 Sample harvesting for analysis

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Kong et al., 2023. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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