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BRAIN Publication-derived

Alzheimer's Disease 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 (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

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

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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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