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BRAIN Publication-derived

COVID-19 Cortical Assembloids

Full SOP with Academic or a plan

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

Authors: 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 Duration: 71 days 4 phases Cell source: 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

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

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Attribution

This SOP was written by Organthis based on the published method in Kong et al., 2023. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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