Skip to content
Back to browse
INTESTINAL Publication-derived

Intestinal Organoids for Drug Screening (2018)

Full SOP with Academic or a plan

Source Vlachogiannis et al. · The Institute of Cancer Research and Royal Marsden NHS Trust, London, UK · 10.1126/science.aao2774

Authors: Vlachogiannis G, Hedayat S, Vatsiou A, Jamin Y, Fernandez-Mateos J, Khan K, Lampis A, Ber K, Romanber J, Pietrber P, Bber JD, Sadanandam A, Cunningham D, Valeri N Duration: 70 days 10 phases Cell source: Patient-Derived (Metastatic CRC Biopsies from Clinical Trials)

Abstract

First prospective clinical study demonstrating that patient-derived tumor organoids from metastatic GI cancer biopsies can predict clinical drug response. Organoid sensitivity testing achieved 100% sensitivity and 93% specificity compared to actual patient outcomes, establishing organoids as viable tools for precision oncology.

Cell source
Patient-Derived (Metastatic CRC Biopsies from Clinical Trials)
Application
Precision Oncology

Protocol overview

43 steps across 10 phases

Phase 1 · Tumor Tissue Acquisition and Processing Day 0
  1. a Patient Enrollment and Biopsy Collection
  2. b Histological Assessment and Sample Preparation
Phase 2 · Patient-Derived Organoid (PDO) Establishment Day 1-14
  1. a Tissue Dissociation and Cell Isolation
  2. b Seeding in 3D Culture System
  3. c PDO Culture Maintenance
  4. d Passaging and Expansion
Phase 3 · Histopathological and Immunohistochemical Characterization Day 7-21
  1. a Specimen Preparation and H&E Staining
  2. b Immunohistochemistry (IHC) for Diagnostic Markers
  3. c Chromogenic In Situ Hybridization (CISH) for Gene Amplifications
  4. d Immunostaining for Biomarker Expression
Phase 4 · Molecular Profiling (NGS and Gene Expression) Day 7-28
  1. a DNA Extraction and Targeted NGS Panel
  2. b Mutational Analysis and Copy Number Variation (CNA) Detection
  3. c Whole-Genome Sequencing (WGS) Validation
  4. d Digital-Droplet PCR (ddPCR) Validation of Key CNAs
  5. e RNA Extraction and Gene Expression Profiling (RNA-Seq)
Phase 5 · High-Throughput 3D Drug Screening Day 21-35
  1. a Organoid Preparation for Drug Screening
  2. b Seeding in 3D Drug Screening Format
  3. c Drug Library Preparation
  4. d Drug Dosing and Treatment
  5. e Cell Viability Assessment (GI50 Determination)
  6. f Hit Validation at Lower Drug Concentrations
Phase 6 · Signaling Pathway Analysis and Mechanism of Action Day 7-14
  1. a Organoid Treatment with Pathway Inhibitors
  2. b Protein Extraction from Treated Organoids
  3. c Western Blot Analysis of Pathway Activation
  4. d Cell Cycle and Apoptosis Assessment
  5. e DNA Damage Assessment
Phase 7 · Establishment of PDO-Derived Orthotopic Xenografts (PDO-Xenografts) Day 28-35
  1. a PDO Preparation for Xenograft Implantation
  2. b Mouse Strain Selection and Xenograft Site
  3. c Surgical Implantation
  4. d Tumor Engraftment Monitoring
Phase 8 · PDO-Xenograft In Vivo Drug Response Studies Day 42-70
  1. a Randomization and Treatment Initiation
  2. b Serial Tumor Monitoring During Treatment
  3. c Functional Imaging (DCE-MRI and Susceptibility-Contrast MRI)
  4. d Survival Monitoring (Long-Term Studies)
Phase 9 · Histopathological Analysis of PDO-Xenografts Day 70 (at endpoint)
  1. a Tumor Harvest and Fixation
  2. b H&E Staining and Morphological Assessment
  3. c Immunohistochemical Assessment of Vasculature
  4. d Assessment of Histopathological Growth Patterns (HGPs)
  5. e Necrosis Quantification
Phase 10 · Correlation of PDO and Clinical Response Data Day 70+
  1. a Compilation of Clinical Response Data
  2. b Matching PDO/Xenograft Responses to Patient Clinical Responses
  3. c Sensitivity, Specificity, and Predictive Value Calculation
  4. d Mechanistic Investigation of Discordant Cases

Full SOP

Full SOPs need an account and a plan

Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.

Create an account

Already registered? Log in

Attribution

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

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence