Cancer invasion and metastasis are challenging to study in vivo since they occur deep inside the body over extended time periods. Organotypic 3D culture of fresh tumor tissue enables convenient real-time imaging, genetic and microenvironmental manipulation and molecular analysis. Here, we provide detailed protocols to isolate and culture heterogenous organoids from murine and human primary and metastatic site tumors. The time required to isolate organoids can vary based on the tissue and organ type but typically takes <7 h. We describe a suite of assays that model specific aspects of metastasis, including proliferation, survival, invasion, dissemination and colony formation. We also specify comprehensive protocols for downstream applications of organotypic cultures that will allow users to (i) test the role of specific genes in regulating various cellular processes, (ii) distinguish the contributions of several microenvironmental factors and (iii) test the effects of novel therapeutics.

癌症侵袭和转移在体内研究具有挑战性，因为它们会在很长一段时间内发生在身体深处。新鲜肿瘤组织的器官型 3D 培养可以方便地进行实时成像、遗传和微环境操作以及分子分析。在这里，我们提供了从小鼠和人类原发性和转移性部位肿瘤中分离和培养异质类器官的详细方案。分离有机物所需的时间可能因组织和器官类型而异，但通常需要 <7 小时。我们描述了一套模拟转移特定方面的分析，包括增殖、存活、入侵、传播和集落形成。