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BRAIN Publication-derived

Brain Organoids for Biobanking

Full SOP with Academic or a plan

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

Authors: 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 Duration: 14 days 5 phases Cell source: 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

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

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Attribution

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

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