Skip to content
← Back to browse
BRAIN Publication-derived

ELAVL4, splicing, and glutamatergic dysfunction precede neuron loss in MAPT mutation cerebral organoids

Source Bowles et al., 2021 · Ronald M. Loeb Center for Alzheimer's Disease, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai · 10.1016/j.cell.2021.07.003

👤 Kathryn R. Bowles, M. Catarina Silva, Kristen Whitney, Taylor Bertucci, Joshua E. Berlind, Jesse D. Lai, Jacob C. Garza, Nathan C. Boles, Sidhartha Mahali, Kevin H. Strang, Jacob A. Marsh, Cynthia Chen, Derian A. Pugh, Yiyuan Liu, Ronald E. Gordon, Susan K. Goderie, Rebecca Chowdhury, Steven Lotz, Keith Lane, John F. Crary, Stephen J. Haggarty, Celeste M. Karch, Justin K. Ichida, Alison M. Goate, Sally Temple ⏱ 180 days 📋 4 phases 🧫 Patient-Derived iPSC (tau-V337M frontotemporal dementia)

Abstract

This protocol describes the generation and characterization of iPSC-derived cerebral organoids expressing the tau-V337M mutation causing frontotemporal dementia. The organoids recapitulate aspects of tauopathy including selective loss of glutamatergic neurons, autophagy-lysosomal dysfunction, tau accumulation, and aberrant splicing driven by dysregulated ELAVL4 expression, providing a platform for identifying early pathological events and testing therapeutic interventions.

Cell source
Patient-Derived iPSC (tau-V337M frontotemporal dementia)
Application
Disease modeling

Protocol overview

15 steps across 4 phases

iPSC Maintenance and Spheroid Formation Day -∞ to Day 0
  1. 1 iPSC Culture Maintenance
  2. 2 Single Cell Dissociation
  3. 3 Cell Washing and Counting
  4. 4 AggreWell Plate Preparation
  5. 5 Spheroid Formation
Neural Induction and Early Differentiation Day 0 to Day 15
  1. 6 Transfer Spheroids to Differentiation Medium
  2. 7 Initial Dorsal Forebrain Patterning
  3. 8 Transition to Neural Medium
  4. 9 Separate Fused Organoids
Neuronal Differentiation and Maturation Day 15 to Day 43
  1. 10 BDNF and NT3 Supplementation
  2. 11 Long-term Maturation
  3. 12 Continue Organoid Separation
Organoid Analysis and Characterization (2-6 months) Day 60 to Day 180
  1. 13 Timepoint Harvest at 2 Months
  2. 14 Timepoint Harvest at 4 Months
  3. 15 Timepoint Harvest at 6 Months

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 Bowles et al., 2021. 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