Fig. 4

HRCA-PDO-based high-throughput/content screening platform has been established for colorectal adenoma drug discovery. A Workflow of high-throughput/content drug screening. HRCA-PDOs from the established biobank were passaged and plated in 96-well plates for 5-day culture, then treated with candidate drugs for four days before staining with PI (red), Hoechst (blue) and Calcein-AM (green), followed by high-content imaging. B Heatmap analysis of the normalized PI/Hoechst ratio in organoids after drug treatment among three HRCA-PDO lines derived from three different patients. Colors range from blue (low mortality) to red (high mortality). P, patient. Thirteen top hits were highlighted with yellow circles. C Bright and fluorescent field imaging illustrated suppressive effect of indicated drugs (artemisinin, berberine, dipyridamole, metformin, and doxazosin) compared with control on HRCA-PDOs. Scale bar, 200 μm. D The normalized surface area of organoids in control and drug-treated groups were calculated and compared. All data were presented as dot plot with mean ± SD (n = 4) and subjected to one-way ANOVA followed by the Bonferroni post hoc test. in D. * indicates P < 0.05 versus control group