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Methods Collections

Prostate Organoid Research Techniques and Methods: Three-Dimensional (3D) Culture, Spin-down, Dimpled Parafilm and Fusion Method

Collection Overview

Organoids are three-dimensional (3D) ex vivo cellular structures that are believed to recapitulate and maintain the in vivo conditions of cellular viability and responsiveness to therapeutic drugs. Matrigel or collagen have been utilized as an extracellular matrix to enable patient-derived cells to form 3D/organoids including cyst-like structures and spheroids. 3D cultured organoids are considered to be excellent models to understand the mechanisms of tumorigenesis and to determine the effects of anticancer agents. Therefore, methods have been developed for a range of cancers such as brain, breast, lung and prostate, among others. Prostate, however, is somewhat unique in that current methods for culturing organoids have a lower success rate than for other cancers. Difficulty in establishing prostate organoid cultures may be driven in part by tumorigenic heterogeneity. Most prostate organoids are derived from patient-derived xenografts (PDXs) and are genomically representative of the patient’s prostate cancer. Therefore, PDX organoids models are invaluable for studying down-stream signaling of target molecules and the effects of anti-cancer agent treatment. Several groups have developed modified protocols that are optimized for their experimental objectives. The goal of this collection is to (1) provide several examples of optimized culture condition for patient-derived prostate cancer 3D organoids, (2) introduce follow-up methodologies for single cell methods such as RNA-FISH and flow cytometry, (3) share meticulous details of immunohistochemistry (IHC)/ immunofluorescent (IF) technique specific for organoids and (4) differential criteria to determine the effects of drug treatment on organoids. 

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