Stem Cell Research & Therapy (May 2022)

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

  • Xiaomeng Li,
  • Guoxiang Fu,
  • Long Zhang,
  • Ruoyu 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

DOI
https://doi.org/10.1186/s13287-022-02902-3
Journal volume & issue
Vol. 13, no. 1
pp. 1 – 14

Abstract

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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. Graphical Abstract

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