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BRAIN Publication-derived

Generation of Cortical-Blood Vessel Assembloids (fCBOs) and SARS-CoV-2 Infection Protocol

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

Abstract

This protocol describes the generation of fused cortical-blood vessel assembloids (fCBOs) by combining cortical organoids with blood vessel organoids to recapitulate Alzheimer's disease and COVID-19 pathology. The fCBOs exhibit increased glial populations, functional vasculature, and susceptibility to SARS-CoV-2 infection, enabling investigation of neuroinflammation and neurodegenerative phenotypes including β-amyloid plaques and hyperphosphorylated tau.

Cell source
Human iPSC
Application
Disease modeling

Protocol overview

18 steps across 4 phases

Cortical Organoid (CO) Generation Days 0-43+
  1. 1 Embryoid body formation
  2. 2 Dual-SMAD inhibition phase
  3. 3 Neural differentiation medium application
  4. 4 Maturation phase with neurotrophic factors
  5. 5 Late maturation
Blood Vessel Organoid (BVO) Generation Days 0-14
  1. 1 Aggregate formation with mesodermal induction
  2. 2 Vascular specification phase
  3. 3 Embedding in extracellular matrix
  4. 4 Blood vessel network extraction
  5. 5 BVO self-assembly
Fusion of Cortical and Blood Vessel Organoids (fCBO Generation) Days 25-71 of cortical differentiation
  1. 1 Organoid co-culture setup
  2. 2 Initial fusion phase
  3. 3 Transfer to 24-well plate with shaking
  4. 4 Transfer to 60 mm dish
  5. 5 Continued maturation
SARS-CoV-2 Infection Day 57 (post-fusion day 32) onwards
  1. 1 Organoid preparation for infection
  2. 2 Viral infection
  3. 3 Incubation and sampling

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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