Skip to content
← Back to browse
BRAIN Publication-derived

Modeling amyloid beta and tau pathology in human cerebral organoids

Source Gonzalez et al., 2018 · University of Texas Medical School at Houston, Department of Neurology, Mitchell Center for Alzheimer's Disease and Related Brain Disorders · 10.1038/s41380-018-0229-8

👤 Cesar Gonzalez, Enrique Armijo, Javiera Bravo-Alegria, Andrea Becerra-Calixto, Charles E. Mays, Claudio Soto ⏱ 110 days 📋 6 phases 🧫 Human iPSC (familial Alzheimer's disease and Down syndrome patient-derived; healthy control iPSC)

Abstract

This protocol describes the generation of cerebral organoids (COs) from human iPSCs derived from Alzheimer's disease (AD) and Down syndrome patients to model AD pathology in a 3D cortical-like tissue structure. The COs spontaneously develop amyloid-beta plaques and tau neurofibrillary tangles characteristic of AD, providing a human cellular model for studying disease mechanisms and screening therapeutic candidates.

Cell source
Human iPSC (familial Alzheimer's disease and Down syndrome patient-derived; healthy control iPSC)
Application
Disease modeling

Protocol overview

31 steps across 6 phases

Fibroblast Culture and iPSC Generation Days 0–30
  1. 1 Culture of human dermal fibroblasts
  2. 2 Prepare lentiviral plasmids
  3. 3 Produce lentiviral particles
  4. 4 Transduce fibroblasts with lentivirus
  5. 5 Culture reprogrammed cells on MEF feeder layers
  6. 6 Select and expand iPSC colonies
iPSC Characterization Days 0–30+ (parallel to expansion)
  1. 1 RT-PCR analysis of pluripotent markers
  2. 2 Alkaline phosphatase (AP) staining
  3. 3 Embryoid body assay
Cerebral Organoid Generation and Maturation Days 0–110
  1. 1 Prepare single-cell suspension from iPSCs
  2. 2 Seed cells in ultra-low attachment 96-well plates
  3. 3 Aggregate formation and expansion (Days 0–6)
  4. 4 Neural induction (Day 6)
  5. 5 Embedding in Geltrex and transfer to 6-well plates (Days 11–12)
  6. 6 Maturation and long-term culture (Days 16+)
Sample Preparation and Immunofluorescence Analysis At 30 DIV and 110 DIV
  1. 1 Fix samples for microscopy
  2. 2 Incubate with primary antibodies
  3. 3 Incubate with secondary antibodies and mount
  4. 4 BTA-1 staining for amyloid aggregates
  5. 5 Hematoxylin staining
  6. 6 Gallyas silver staining for neurofibrillary tangles
  7. 7 Image acquisition
Biochemical Analysis: Western Blot and ELISA At 110 DIV
  1. 1 Collect and prepare organoid homogenates
  2. 2 Quantify protein concentration
  3. 3 Western blot sample preparation
  4. 4 Transfer to membrane and blocking
  5. 5 Incubate with primary and secondary antibodies (phospho-tau detection)
  6. 6 Develop and visualize blots
  7. 7 ELISA sample preparation: Fractionation of Aβ and tau
  8. 8 ELISA for Aβ and tau quantification
Statistical Analysis During data analysis
  1. 1 Data analysis for Aβ and tau aggregates

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

This SOP was authored by Organthis based on the published method in Gonzalez et al., 2018. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence