Engineering microcavity arrays as a niche for high-throughput and high-content analysis of tumoroids: A detailed methodology.

Tumoroids, which replicate patient tumor genetics and drug responses, have revolutionized oncology by enabling personalized treatment development and reducing reliance on animal testing. Despite remarkable advances in reproducing ever more sophisticated biology in advanced organoid platforms, there remain foundational challenges in standardizing and scaling-up tumour organoid development. Moreover, recreating key aspects of the tumour niche, namely its biomechanical properties is still overlooked in drug screening. To address these shortcomings, we introduce a technology for high-throughput derivation and high-content screening of tumoroids. Our technology relies on biofabrication of microengineered cell culture devices, which are scalable and tailored for standard automated lab-setups and real-time analysis of thousands of individual tumoroids trapped in microcavity arrays, within a polymer-hydrogel substrate. We validated our system using patient-derived urothelial cancer tumoroids, as they represent a good example of one of the most frequent cancer types with a debilitating clinical entity and a high financial burden related to the disease recurrency. Our scalable-culture technology should open new avenues toward more patient-tailored treatments or even new drugs targeting the extracellular matrix. Ultimately, the economic burden of the disease can be decreased by increasing treatment efficiency through larger, faster and more reliable screening.