A Feasibility Test of the Disposable Drug Daughter Plate Combined with 384-Pillar Plate for Patient-Derived Organoid-Based High Throughput Screening

A disposable daughter plate is developed for robotic organoid-based high-throughput screening (HTS). Previously, daughter drug plates are prepared using mother drug plates in conventional HTS. During each screening, the mother drug plate is thawed, a certain amount of the drug is transferred to the daughter plate, and frozen again. Drug manufacturing involves drug transfer processes that may result in drug damage owing to redissolution. Thus, disposable daughter plates are created by dispensing small drug spots into the well plates and freezing them. The cells at the Matrigel dorms of the pillar plate are easily exposed to the drugs by immersing the pillars. Unlike previously reported similar methods, it is verified that HTS-based drug response analysis is possible by applying the proposed disposable daughter plate to breast cancer patient-derived organoids (PDOs). And it is also verified the long-term stability of disposable daughter plates (up to 3 months) in 3D cell culture system using patient-derived cells (PDO). The drug response results are quantified using normalized area under curve values, while the experimental reproducibility of the disposable daughter plates is evaluated using the coefficient of variation and a simple linear regression test. Thus, the proposed disposable daughter plate allows reproducible drug efficacy analysis. A disposable daughter plate is proposed that improves the limitations of conventional drug treatment methods. The proposed disposable daughter plate can be stored for a long time and is used for single use, solving the problem of cross-contamination. The feasibility of the disposable daughter plate is verified through reproducibility analysis of high-throughput drug screening. image