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BRAIN Publication-derived

Individualized Patient Tumor Organoid (IPTO) Model for Brain Cancer Culture and Drug Response Prediction

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

👤 Tianping Peng, Xiujian Ma, Wei Hua, Yonghe Wu, Hai-Kun Liu, Ying Mao ⏱ 20 days 📋 5 phases 🧫 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

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

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Peng et al., 2025. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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