Fig. 2
Experimental procedures of HTS using lung cancer PDOs (A) Tumor tissue and malignant pleural effusion (MPE) samples derived from patients with lung cancer. (B) Preparation step for isolating cancer cells from patient-derived lung cancer tumor tissues and MPEs. (C) The pretreated patient-derived lung cancer samples filtered using a 40 μm strainer. (D) Patient-derived lung cancer cells mixed with the extracellular matrix (ECM) of hydrogel components. (E) Cancer cell and Matrigel mixtures dispensed on the surface of 384-pillar plate by ASFA™ Spotter DN. (F) The drug exposed by combining the 384-pillar plate, in which the cells have been dispensed, with the 384-well plates containing the fresh culture medium and drugs. (G) 3D live cell staining by replacing the 384-well plate with one filled with live cell-staining dye; images were taken after live cell staining. (H) Quantifying the percentage of live cells by evaluating scanned images and calculating the cell growth rate. (I) Cell viability measurement through CellTiter-Glo reagent treatment and luminescence value readout; drug sensitivity analysis based on AUC index from dose-response curve (DRC). (J) CODRP index predicted drug efficacy by considering the cell growth rate and cancer diagnosed stage in the AUC index. (K) Immunohistochemistry (IHC) analysis using patient-derived lung cancer tissues and cultured lung cancer organoid samples