High-throughput colorectal cancer organoid-on-a-chip for single-organoid-analysis-based personalized drug evaluation

Patient-derived colorectal cancer organoids (CRCOs) are increasingly recognized for their ability to preserve genetic, transcriptional, histological, and drug-responsive features of the original colorectal tumors, offering promise in precision medicine and drug development. While the conventional Matrigel dome culture (DOME) approach has been instrumental in enabling valuable investigations, it has limitations—most notably considerable variation in organoid size and drug sensitivity profiles. In this study, we developed an organoid-on-a-chip (CHIP) for forming uniform organoids and improving precision drug evaluation. Specifically, we fabricated the CHIP using three-dimensional (3D)-printing and micro-molding techniques. Using this CHIP, 19 uniform CRCOs can be generated concurrently in a single well of a 96-well plate within three days, each forming from a minimal seeding density of only 100 cells. Furthermore, the CHIP facilitates in situ imaging and continuous monitoring of individual organoids, supporting label-free, morphometric analysis-based drug screening. Organoids cultured on the CHIP show significantly improved consistency, uniformity, and reproducibility over the DOME method. Our CHIP organoids are functionally identical to conventional DOME organoids, but exhibit greater reliability in drug screening. These results support the CHIP’s potential for high-throughput assays and predictive drug screening models.