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BRAIN Publication-derived

Frontotemporal Dementia Cerebral Organoids for Disease Modelling

Full SOP with Academic or a plan

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

Authors: 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 Duration: 180 days 4 phases Cell source: 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

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

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Attribution

This SOP was written by Organthis based on the published method in Bowles et al., 2021. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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