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BRAIN Publication-derived

Glioblastoma Organoids for Functional Precision Oncology

Source Jacob et al. · University of Pennsylvania, Philadelphia, PA, USA · 10.1016/j.cell.2019.11.036

👤 Jacob F, Salinas RD, Zhang DY, Nguyen PTT, Schnoll JG, Wong SZH, Thokber R, Sheikh S, Saxena D, Prokber S, Bhatt SM, Bhatt AH, Liu D, Bhatt RM, Song H, Ming GL ⏱ 14 days 📋 5 phases 🧫 Patient-Derived Primary GBM Tissue (Biobank, >70 Patients)

Abstract

Jacob et al. established the largest GBM organoid biobank from over 70 patient tumors, generating organoids within 2 weeks that retain the mutational landscape and native immune microenvironment including T cells and macrophages. This Cell 2020 platform enables functional precision oncology including immunotherapy and CAR-T cell testing on patient-specific GBM organoids.

Cell source
Patient-Derived Primary GBM Tissue (Biobank, >70 Patients)
Application
Functional Precision Oncology

Protocol overview

21 steps across 5 phases

Tissue Collection and Preparation Day 0
  1. 1 Surgical specimen transfer to pathology
  2. 2 Transition to culture medium
  3. 3 Tissue mincing and debris removal
  4. 4 Red blood cell lysis
  5. 5 Aliquoting for analysis and culture
GBO Establishment and Initial Culture Day 0–14
  1. 1 Transition to GBO culture medium
  2. 2 Orbital shaker culture initiation
  3. 3 Monitor organoid formation (Days 1–14)
  4. 4 Medium change (if needed)
GBO Passaging and Maintenance (Week 2+) Day 14 onwards
  1. 1 Prepare organoids for passaging
  2. 2 Cut organoids into smaller pieces
  3. 3 Transfer to fresh culture plate
  4. 4 Routine medium changes
GBO Characterization (Optional at 2 and 4 weeks) Week 2–4
  1. 1 Harvest samples for immunohistology
  2. 2 Cryoprotection and freezing
  3. 3 Harvest samples for RNA sequencing
  4. 4 Hypoxia gradient detection (optional)
GBO Biobanking and Cryopreservation Week 2–4 (or at desired passage)
  1. 1 Prepare GBOs for freezing
  2. 2 Slow cooling (optional controlled-rate freezing)
  3. 3 Long-term storage in liquid nitrogen
  4. 4 Recovery from cryopreservation

Full SOP

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Attribution

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

This wording is awaiting legal review.

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