An Automation Workflow for High‐Throughput Manufacturing and Analysis of Scaffold‐Supported 3D Tissue Arrays

Abstract

AbstractPatient‐derived organoids have emerged as a useful tool to model tumour heterogeneity. Scaling these complex culture models while enabling stratified analysis of different cellular sub‐populations, however, remains a challenge. One strategy to enable higher throughput organoid cultures is the scaffold‐supported platform for organoid‐based tissues (SPOT). SPOT allows the generation of flat, thin, and dimensionally‐defined microtissues in both 96‐ and 384‐well plate footprints that are compatible with longitudinal image‐based readouts. SPOT is currently manufactured manually, however, limiting scalability. In this study, an automation approach to engineer tumour‐mimetic 3D microtissues in SPOT using a liquid handler is optimized and comparable within‐ and between‐sample variation to standard manual manufacturing is shown. Further, a liquid handler‐supported cell extraction protocol to support single‐cell‐based end‐point analysis using high‐throughput flow cytometry and multiplexed cytometry by time of flight is developed. As a proof‐of‐value demonstration, 3D complex tissues containing different proportions of tumour and stromal cells are generated to probe the reciprocal impact of co‐culture. It is also demonstrated that primary patient‐derived organoids can be incorporated into the pipeline to capture patient‐level tumour heterogeneity. It is envisioned that this automated 96/384‐SPOT workflow will provide opportunities for future applications in high‐throughput screening for novel personalized therapeutic targets.