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BRAIN Publication-derived

Glioblastoma Cerebral Organoids for Drug Screening

Full SOP with Academic or a plan

Source Peng et al., 2025 · German Cancer Research Center (DKFZ), Heidelberg; Fudan University, Shanghai · 10.1016/j.stem.2025.01.002

Authors: Tianping Peng, Xiujian Ma, Wei Hua, Yonghe Wu, Hai-Kun Liu, Ying Mao Duration: 20 days 5 phases Cell source: Patient-Derived Tumor Explant (Primary and Metastatic Brain Cancers)

Abstract

This protocol describes the generation and culture of individualized patient tumor organoids (IPTOs) from primary and metastatic brain cancers using patient-derived tumor explants co-cultured with iPSC-derived cerebral organoids. IPTOs maintain cellular heterogeneity, tumor microenvironment, and molecular features of original patient tumors, enabling prediction of patient-specific drug responses including resistance mechanisms.

Cell source
Patient-Derived Tumor Explant (Primary and Metastatic Brain Cancers)
Application
Disease modeling and drug response prediction

Protocol overview

27 steps across 5 phases

Phase 1 · iPSC Culture and Cerebral Organoid Generation Days 0-20+
  1. a iPSC Culture Maintenance
  2. b Dissociate iPSCs to Single Cells
  3. c Embryoid Body Formation
  4. d Organoid Embedding in Matrigel
  5. e Differentiation Phase (Days 13-19)
  6. f Differentiation Phase with Vitamin A (Day 20+)
Phase 2 · Tumor Tissue Acquisition and Processing Day 0-1 (upon surgical resection)
  1. a Tissue Collection and Transport
  2. b Wash and Dissect Tumor Fragments
  3. c Prepare Tumor Fragments for Multiple Uses
Phase 3 · Co-culture of Tumor Explants with Cerebral Organoids (IPTO Generation) Day 1-14+ (typically analyzed at 2 weeks)
  1. a Prepare Cerebral Organoids
  2. b Insert Tumor Fragments into Organoid Pocket
  3. c Embed in Matrigel
  4. d Incubate for Matrigel Gelation
  5. e Transfer IPTOs to Culture Plates
  6. f Stationary Culture Overnight
  7. g Transfer to Orbital Shaker
  8. h Routine Medium Changes
  9. i Monitor IPTO Growth (2-14 weeks typical)
Phase 4 · Lentiviral Luciferase Labeling for Bioluminescence Imaging Concurrent with IPTO generation (Day 1-2)
  1. a Prepare Lentiviral Particles
  2. b Infect Tumor Tissues with Lentivirus
  3. c Wash Virus from Tissue
  4. d Proceed with IPTO Assembly
Phase 5 · Drug Treatment and Bioluminescence Imaging Days 7-14 (typical treatment window post-IPTO generation)
  1. a Prepare Drug Solutions
  2. b Add Drug to IPTO Cultures
  3. c Continue Culture with Drug
  4. d Perform Bioluminescence Imaging (BLI)
  5. e Calculate Drug Response

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Attribution

This SOP was written by Organthis based on the published method in Peng et al., 2025. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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