Figure 2.

Establishment and characterization of patient-derived PCa organoids and corresponding tissue. (A) Representative bright-field image showing established PCa organoids (G1) grown in culture. Scale bar, 100 µm. Mean, minimum and maximum (B) OFC and (C) diameter of patient-derived PCa organoids (n=10). (D) Immunofluorescent images of organoids [patient 22; grade group 5; Gleason score, 9 (5+4)] stained with prostate lineage epithelial markers CK8 and CK5 revealing the presence of both prostate epithelial lineages in the established organoid cultures, with organoids expressing luminal- or basal-only or luminal and basal double-positive cells. (E) Immunofluorescent images of organoids and corresponding tissue stained with prostate lineage epithelial markers CK8 and CK5 [patient 8; grade group 1; Gleason score, 6 (3+3)] and mesenchymal marker VIM [patient 25; grade group 3; Gleason score 7 (4+3)]. The nuclei were stained with anti-fade Fluorogel II with DAPI. Representative confocal microscopy images were acquired using a Zeiss LSM 710 laser scanning confocal microscope and processed using Carl Zeiss ZEN 2013 image software. (F) Immunohistochemistry images of organoids and corresponding tissue [patient 7; grade group 2; Gleason score 7 (3+4)] stained with H&E and prostate lineage epithelial markers p63, AR and PSA. Scale bar, 50 µm. Representative microscopy images were acquired using an Olympus CX41 light microscope (×10 magnification). Inset magnification, ×2.5. (G) RNA-seq gene expression principal component analysis plot of PCa organoids and corresponding tissue for two patients [patient 1, grade group 1/Gleason score 6 (3+3); patient 2, grade group 2/Gleason score 7 (3+4)]. PCa, prostate cancer; G, generation; OFC, organoid formation count; CK, cytokeratin; VIM, vimentin; H&E, hematoxylin and eosin; AR, androgen receptor; PSA, prostate-specific antigen.