Adult stem cell–based organoid technology is a versatile tool for the generation and long-term maintenance of near-native 3D epithelial tissues in vitro. The generation of cancer organoids from primary patient material enables a range of therapeutic agents to be tested in the resulting organoid cultures. Patient-derived cancer organoids therefore hold great promise for personalized medicine. Here, we provide an overview of the protocols used by different groups to establish organoids from various epithelial tissues and cancers, plus the different protocols subsequently used to test the in vitro therapy sensitivity of these patient-derived organoids. We also provide an in-depth protocol for the generation of head and neck squamous cell carcinoma organoids and their subsequent use in semi-automated therapy screens. Establishment of organoids and subsequent screening can be performed within 3 months, although this timeline is highly dependent on a.o. starting material and the number of therapies tested. The protocol provided may serve as a reference to successfully establish organoids from other cancer types and perform drug screenings thereof.

基于成体干细胞的类器官技术是一种多功能工具，用于在体外生成和长期维持近天然 3D 上皮组织。从主要患者材料中生成癌症类器官使得可以在所得类器官培养物中测试一系列治疗剂。因此，源自患者的癌症类器官在个性化医疗方面具有广阔的前景。在这里，我们概述了不同小组用于从各种上皮组织和癌症中建立类器官的方案，以及随后用于测试这些源自患者的类器官的体外治疗敏感性的不同方案。我们还提供了用于生成头颈鳞状细胞癌类器官及其随后在半自动治疗筛选中的使用的深入方案。类器官的建立和随后的筛选可以在 3 个月内完成，尽管这个时间表高度依赖于起始材料和测试的疗法的数量。所提供的方案可以作为成功建立其他癌症类型的类器官并对其进行药物筛选的参考。