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
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.
Protocol overview
18 steps across 4 phases
- 1 Embryoid body formation
- 2 Dual-SMAD inhibition phase
- 3 Neural differentiation medium application
- 4 Maturation phase with neurotrophic factors
- 5 Late maturation
- 1 Aggregate formation with mesodermal induction
- 2 Vascular specification phase
- 3 Embedding in extracellular matrix
- 4 Blood vessel network extraction
- 5 BVO self-assembly
- 1 Organoid co-culture setup
- 2 Initial fusion phase
- 3 Transfer to 24-well plate with shaking
- 4 Transfer to 60 mm dish
- 5 Continued maturation
- 1 Organoid preparation for infection
- 2 Viral infection
- 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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