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OTHER Publication-derived

Establishment and Derivation of Patient-Derived Tumor Organoids for Cancer Research and Drug Screening

Source Drost et al., 2018 · Princess Máxima Centre for Paediatric Oncology and Hubrecht Institute, Utrecht, Netherlands · 10.1038/s41586-018-0024-3

👤 Jarno Drost, Hans Clevers ⏱ 21 days 📋 8 phases 🧫 Patient-Derived Tumor Tissue (multiple cancer types)

Abstract

This protocol describes the establishment of long-term organoid cultures from primary patient-derived tumor and matching healthy tissue samples. Tumor organoids are derived by embedding tissue samples in Matrigel and culturing under serum-free conditions with tissue-specific growth factors. These organoid cultures maintain genetic and phenotypic features of the original tumors and enable rapid drug screening and personalized medicine approaches.

Cell source
Patient-Derived Tumor Tissue (multiple cancer types)
Application
Disease modeling, Drug screening, Personalized medicine

Protocol overview

21 steps across 8 phases

Tissue Sample Preparation and Processing Day 0
  1. 1 Obtain primary tumor and matched normal tissue biopsy samples
  2. 2 Mechanically dissociate tissue samples
Organoid Initiation in Matrigel Day 0–1
  1. 3 Prepare Matrigel-containing culture wells
  2. 4 Apply selective culture conditions if needed
Standard Culture Medium Supplementation Day 1 onwards
  1. 5 Prepare serum-free culture medium
  2. 6 Add p38 MAPK inhibitor and other tissue-specific factors if required
Long-Term Culture Maintenance and Expansion Day 7 onwards
  1. 7 Replace culture medium regularly
  2. 8 Perform organoid passaging and re-plating
  3. 9 Cryopreserve organoids for biobanking
Genetic and Phenotypic Characterization Upon organoid establishment and periodically during culture
  1. 10 Verify tumor origin with mutation analysis
  2. 11 Assess phenotypic features and biomarker expression
  3. 12 Perform proteomic and transcriptomic profiling
Drug Screening on Patient-Derived Organoids Upon organoid establishment, typically day 14–21 of culture
  1. 13 Prepare organoid cultures for drug screening
  2. 14 Apply drug compounds and drug libraries
  3. 15 Assess drug-induced phenotypes and cell viability
  4. 16 Validate hepatotoxicity using healthy hepatocyte organoids
Xenotransplantation Validation (Optional) Upon completion of in vitro drug screening
  1. 17 Transplant organoids into immunodeficient mice
  2. 18 Perform in vivo drug treatment studies
Mutational Process Studies (Optional) Upon clonal organoid establishment
  1. 19 Establish clonal organoid cultures
  2. 20 Perform whole genome sequencing on clonal organoids
  3. 21 Study effects of chemotherapy or radiation on mutation profiles

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Drost et al., 2018. 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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