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BRAIN Publication-derived

Lower-Grade Glioma Brain Tumouroids

Full SOP with Academic or a plan

Source Abdullah et al., 2022 · University of Texas Southwestern Medical Center · 10.1093/neuonc/noab273

Authors: Kalil G. Abdullah, Cylaina E. Bird, Joseph D. Buehler, Lauren C. Gattie, Milan R. Savani, Alex C. Sternisha, Yi Xiao, Michael M. Levitt, William H. Hicks, Wenhao Li, Denise M.O. Ramirez, Toral Patel, Tomas Garzon-Muvdi, Samuel Barnett, Gao Zhang, David M. Ashley, Kimmo J. Hatanpaa, Timothy E. Richardson, Samuel K. McBrayer Duration: 28 days 4 phases Cell source: Patient-Derived Glioma Tissue (Primary resection samples)

Abstract

This protocol describes an optimized method for establishing patient-derived organoid models from lower-grade glioma (LGG) primary tumor tissue samples using physiologic 5% oxygen conditions. The resulting organoids recapitulate parental tumor characteristics including cytoarchitecture, cellular heterogeneity, proliferative capacity, and genomic alterations, providing a faithful in vitro platform for preclinical studies of LGG biology and treatment response assessment.

Cell source
Patient-Derived Glioma Tissue (Primary resection samples)
Application
Disease modeling and drug screening

Protocol overview

15 steps across 4 phases

Phase 1 · Tissue Processing and Initial Culture Preparation Day 0-1
  1. a Tumor tissue collection and transport
  2. b Red blood cell lysis
  3. c Tissue washing
  4. d Tissue dissection
  5. e Organoid plating
Phase 2 · Organoid Culture and Maintenance Day 1-28+ (minimum 4 weeks)
  1. a Incubation setup
  2. b Medium refreshment
  3. c Culture duration and monitoring
Phase 3 · Cryopreservation and Biobanking Variable (after minimum 4 weeks in culture)
  1. a Organoid grouping for freezing
  2. b DMSO addition and cryotube preparation
  3. c Controlled-rate freezing
  4. d Long-term storage in liquid nitrogen
  5. e Reanimation after biobanking
Phase 4 · Tissue Fixation and Preparation for Analysis At analysis timepoint (minimum 4 weeks post-explantation)
  1. a Organoid fixation
  2. b Formalin removal and ethanol preservation

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Attribution

This SOP was written by Organthis based on the published method in Abdullah et al., 2022. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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