Department of Anatomy, Cell Biology, and Physiological Sciences, Faculty of Medicine, American University of Beirut, Beirut, Lebanon.
Methods Mol Biol. 2024;2777:135-144. doi: 10.1007/978-1-0716-3730-2_10.
Prostate cancer (PCa) is the second most common malignancy and the fifth leading cause of cancer death in men worldwide. Despite its prevalence, the highly heterogenic PCa has shown difficulty to establish representative cell lines that reflect the diverse phenotypes and different stages of the disease in vitro and hence hard to model in preclinical research. The patient-derived organoid (PDO) technique has emerged as a groundbreaking three-dimensional (3D) tumor modeling platform in cancer research. This versatile assay relies on the unique ability of cancer stem cells (CSCs) to self-organize and differentiate into organ-like mini structures. The PDO culture system allows for the long-term maintenance of cancer cells derived from patient tumor tissues. Moreover, it recapitulates the parental tumor features and serves as a superior preclinical model for in vitro tumor representation and personalized drug screening. Henceforth, PDOs hold great promise in precision medicine for cancer. Herein, we describe the detailed protocol to establish and propagate organoids derived from isolated cell suspensions of PCa patient tissues or cell lines using the 3D semisolid Matrigel™-based hanging-drop method. In addition, we highlight the relevance of PDOs as a tool for evaluating drug efficacy and predicting tumor response in PCa patients.
前列腺癌(PCa)是全球男性中第二大常见恶性肿瘤和第五大癌症死亡原因。尽管它很普遍,但高度异质的 PCa 一直难以建立能够反映疾病不同表型和不同阶段的代表性细胞系,因此难以在临床前研究中建模。患者来源的类器官(PDO)技术已成为癌症研究中开创性的三维(3D)肿瘤建模平台。这种多功能检测依赖于癌症干细胞(CSC)自我组织和分化为类器官样小结构的独特能力。PDO 培养系统允许长期维持源自患者肿瘤组织的癌细胞。此外,它再现了亲本肿瘤的特征,并作为体外肿瘤代表性和个性化药物筛选的优越临床前模型。因此,PDO 在癌症精准医学中具有广阔的前景。本文详细描述了使用基于 3D 半固态基质胶的悬滴法从 PCa 患者组织或细胞系分离的细胞悬液中建立和扩增类器官的方法。此外,我们强调了 PDO 作为评估 PCa 患者药物疗效和预测肿瘤反应的工具的相关性。