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BRAIN Publication-derived

Establishment of patient-derived organoid models of lower-grade glioma

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

👤 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 ⏱ 28 days 📋 4 phases 🧫 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

Tissue Processing and Initial Culture Preparation Day 0–1
  1. 1 Tumor tissue collection and transport
  2. 2 Red blood cell lysis
  3. 3 Tissue washing
  4. 4 Tissue dissection
  5. 5 Organoid plating
Organoid Culture and Maintenance Day 1–28+ (minimum 4 weeks)
  1. 6 Incubation setup
  2. 7 Medium refreshment
  3. 8 Culture duration and monitoring
Cryopreservation and Biobanking Variable (after minimum 4 weeks in culture)
  1. 9 Organoid grouping for freezing
  2. 10 DMSO addition and cryotube preparation
  3. 11 Controlled-rate freezing
  4. 12 Long-term storage in liquid nitrogen
  5. 13 Reanimation after biobanking
Tissue Fixation and Preparation for Analysis At analysis timepoint (minimum 4 weeks post-explantation)
  1. 14 Organoid fixation
  2. 15 Formalin removal and ethanol preservation

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Abdullah et al., 2022. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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