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Assay establishment and validation of a high-throughput organoid-based drug screening platform

2022; BioMed Central; Volume: 13; Issue: 1 Linguagem: Inglês

10.1186/s13287-022-02902-3

1757-6512

Xiaomeng Li, Guoxiang Fu, Long Zhang, Ruo‐Yu Guan, Peiyuan Tang, Jialing Zhang, Xinxin Rao, Shengzhi Chen, Xiaoya Xu, Yi Zhou, Yun Deng, Tao� Lv, Xingfeng He, Shaobo Mo, Peiyuan Mu, Jianjun Gao, Guoqiang Hua,

Cell Image Analysis Techniques

Organoids are three-dimensional structures that closely recapitulate tissue architecture and cellular composition, thereby holding great promise for organoid-based drug screening. Although growing in three-dimensional provides the possibility for organoids to recapitulate main features of corresponding tissues, it makes it incommodious for imaging organoids in two-dimensional and identifying surviving organoids from surrounding dead cells after organoids being treated by irradiation or chemotherapy. Therefore, significant work remains to establish high-quality controls to standardize organoid analyses and make organoid models more reproducible.In this study, the Z-stack imaging technique was used for the imaging of three-dimensional organoids to gather all the organoids' maximum cross sections in one imaging. The combination of live cell staining fluorescent dye Calcein-AM and ImageJ assessment was used to analyze the survival of organoids treated by irradiation or chemotherapy.We have established a novel quantitative high-throughput imaging assay that harnesses the scalability of organoid cultures. Using this assay, we can capture organoid growth over time, measure multiple whole-well organoid readouts, and show the different responses to drug treatments.In summary, combining the Z-stack imaging technique and fluorescent labeling methods, we established an assay for the imaging and analysis of three-dimensional organoids. Our data demonstrated the feasibility of using organoid-based platforms for high-throughput drug screening assays.