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- Title
Assay establishment and validation of a high-throughput organoid-based drug screening platform.
- Authors
Li, Xiaomeng; Fu, Guoxiang; Zhang, Long; Guan, Ruoyu; Tang, Peiyuan; Zhang, Jialing; Rao, Xinxin; Chen, Shengzhi; Xu, Xiaoya; Zhou, Yi; Deng, Yun; Lv, Tao; He, Xingfeng; Mo, Shaobo; Mu, Peiyuan; Gao, Jianjun; Hua, Guoqiang
- Abstract
Background: 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. Methods: 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. Results: 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. Conclusions: 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.
- Subjects
THREE-dimensional imaging; FLUORESCENT dyes; STAINS &; staining (Microscopy); HIGH throughput screening (Drug development)
- Publication
Stem Cell Research & Therapy, 2022, Vol 13, Issue 1, p1
- ISSN
1757-6512
- Publication type
Article
- DOI
10.1186/s13287-022-02902-3