Figure 1 from Human 3D Ovarian Cancer Models Reveal Malignant Cell–Intrinsic and –Extrinsic Factors That Influence CAR T-cell Activity

OvCAR3 cells were sensitive but G164 cells were resistant to CAR T-cell cytotoxicity in monolayer cultures. A, Flow cytometry analysis of the expression of MUC1 and TnMUC1 on monolayer, spheroid, and collagen gel cultures of OvCAR3 and G164 cells. Data plotted as mean ± SD of three repeats. B, Expression of MUC1 and TnMUC1 on OvCAR3+FB and G164+FB collagen gels. Data plotted as mean ± SD of three gels per three replicates. Different FB donors were used for each replicate. Scale bars, 50 μm. C, Schematic representation of CD19, MUC1, and TnMUC1 directed second generation CARs. D, ELISA data showing IFNγ concentration after coculturing CAR T cells with monolayers of T47D, OvCAR3, and G164 cells for 2 days at 1:1 and 1:5 T:E ratios. E, Representative images (left) and quantification (right) of Incucyte killing assay in which monolayers of T47D, OvCAR3, and G164 cells were treated with CAR T cells from one donor at 1:5 T:E ratio. Images shown are 3 days after treatment. Red, dead cells. Scale bars, 400 μm. F, Flow cytometry analysis of the expression of T-cell activation markers HLA-DR (top) and CD25 (bottom) pre and 2 days post coculture with monolayers of T47D, OvCAR3, and G164 cells. D and F, Data plotted as mean ± SD for three CAR T-cell donors. Statistics performed using two-way (D) and one-way ANOVA (F).