Novel therapeutic diiminoquinone exhibits anticancer effects on human colorectal cancer cells in two-dimensional and three-dimensional in vitro models

Figure 8 Establishment and characterization of patient-derived organoids from colon cancer patient 1. A: Representative image of organoids derived from patient 1 stained with hematoxylin and eosin (HE); B: Representative bright-field images of organoids at generation (G)1, G2, and G6. Fresh tumor tissues were enzymatically digested, and single cells were plated in 90% growth factor-reduced Matrigel. G1 organoids were successfully propagated up to G6. Images were visualized by Axiovert inverted microscope at × 5, × 10, and × 20 magnification. Scale bar 100 μm; C: Immunofluorescent images of organoids stained with colon lineage epithelial markers cytokeratin (CK)19 and CK8 and stem cell marker CD44. The nuclei were stained with anti-fade Fluorogel II with 4’,6-diamidino-2-phenylindole (DAPI). Representative confocal microscopy images were acquired using a Zeiss LSM 710 laser scanning confocal microscope. Scale bar 100 μm; D: Representative bright-field images of G2 organoids treated with diiminoquinone (DIQ) (0.5 and 1 μmol/L) or 5-fluorouracil (5FU) (3 μmol/L). Organoid formation count (OFC) and size were calculated, and mean values were reported as mean ± standard error of the mean (^aP < 0.05, ^bP < 0.01, ^cP < 0.001). Images were visualized by Axiovert inverted microscope at × 5 and × 20 magnification. Scale bar 100 μm.