Targeting Immune-Related Molecules in Cancer Therapy: A Comprehensive In Vitro Analysis on Patient-Derived Tumor Models

Figure 5

(a) Apoptosis/necrosis discrimination and (b) coculture of tumor and immune cells. (a) Quantitative analyses of cell death using flow cytometric Yo-Pro-1/PI staining after incubation with test substances for 72 h. For each sample, 10,000 events were measured. Dead cells were defined as early apoptotic (YO-PRO-1⁺), late apoptotic (YO-PRO-1⁺/PI⁺), or necrotic (PI⁺). Given are the % numbers of stained cells after treatment. N = 3 independent experiments, mean ± SD, p < 0.05 versus control. t-test. (b) Tumor and immune cells (PBL) from three healthy donors were coincubated in the presence or absence of test substances for 72 h. Tumor cell quantification was conducted by bead-based flow cytometry as stated in material and methods. Cells cocultured with PBLs but without addition of test substances were used as control. Percentage numbers of residual tumor cells for each donor are shown after 72 h and incubation with test substances. N = 3 PBL donor. HROC, Hansestadt Rostock Colon; CIMP, CpG island methylator phenotype; MSI, microsatellite instability/instable.

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