Abstract Prostate cancer (PCa) is the most commonly diagnosed malignant tumour in men and the second leading cause of cancer-related mortality worldwide. Despite the high long-term survival rates in localised PCa, metastatic disease remains largely incurable even after intensive multimodal therapy. Our limited understanding of the drivers of aggressive behaviour in advanced PCa and the molecular pathways that underlie their failure to respond to various drug therapies underscores the need to study tumours in vitro so that more effective therapies can be evaluated. Tumour-derived organoids represent an advanced in vitro model for studying cancer and are increasingly being used in the development of new therapeutic strategies, serving as valuable tools for advancing precision medicine in oncology. Our goal was to characterise patient-derived organoids from our prostate samples cohort of the FMRP-USP, which accurately model the original patient tumours. Our approach involved establishing longer-term organoid cultures from biopsy samples in Matrigel. For this, we used 9 fresh prostatectomy tissue samples that were processed by mechanical dissection and enzymatic digestion, then filtered and cultured in advanced DMEM/F12 medium with supplements (GlutaMAX, B27, N-acetylcysteine, recombinant EGF, FGF-10 and FGF-2, A-83-01, SB202190, nicotinamide, DHT, PGE2, Y-27632, Noggin, and R-spondin). Nine patients were aged between 50 and 75 years and diagnosed with high-risk PCa with ISUP grade of 4 or 5 (Ethics Committee Number 88216025.3.0000.5440). The organoids were characterized using morphological 3D imaging and histological (H Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 738.
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