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Updated: Apr 11, 2026
Establishment and Maintenance of Patient-derived Prostate Cancer Organoids: A Detailed Experimental Protocol
Published on: November 14, 2025
Protocol for generating and characterizing Matrigel-free prostate cancer patient-derived organoids
Robin Dolgos1, Romuald Parmentier1, Jing Wang1
1Institute of Medical Genetics and Pathology, University Hospital Basel, Basel, Switzerland; Department of Urology, University Hospital Basel, Basel, Switzerland; Department of Biomedicine, University of Basel, University Hospital Basel, Basel, Switzerland.
This study introduces a Matrigel-free protocol for creating prostate cancer patient-derived organoids (PCa PDOs). The method includes sample processing, organoid generation, characterization, and scRNA-seq analysis guidance.
Area of Science:
- Oncology
- Biotechnology
- Genomics
Background:
- Prostate cancer patient-derived organoids (PCa PDOs) are valuable tools for cancer research.
- Current organoid generation protocols often rely on Matrigel, which can introduce variability.
- Standardized protocols are needed for reliable PCa PDO generation and downstream analysis.
Purpose of the Study:
- To present a detailed protocol for generating prostate cancer patient-derived organoids (PCa PDOs) without using Matrigel.
- To provide comprehensive guidance on the entire workflow, from sample handling to bioinformatic analysis.
- To facilitate the use of PCa PDOs for research and the Prostate PDO Single-Cell Atlas (PPScA).
Main Methods:
- Detailed steps for pathological resection and tissue dissociation of prostate cancer samples.
- Procedures for the successful generation, maintenance, and characterization of PCa PDOs in Matrigel-free conditions.
- Methodology for processing PCa PDOs for single-cell RNA sequencing (scRNA-seq) analysis.
Main Results:
- Successful generation of prostate cancer patient-derived organoids (PCa PDOs) using a Matrigel-free protocol.
- Established a robust workflow for organoid maintenance and characterization.
- Provided a framework for scRNA-seq data processing and utilization of the Prostate PDO Single-Cell Atlas (PPScA).
Conclusions:
- The presented Matrigel-free protocol enables reproducible generation of PCa PDOs.
- This protocol supports advanced research applications, including scRNA-seq analysis.
- The Prostate PDO Single-Cell Atlas (PPScA) serves as a valuable resource for researchers studying prostate cancer heterogeneity.
