Figure 7 from Human 3D Ovarian Cancer Models Reveal Malignant Cell–Intrinsic and –Extrinsic Factors That Influence CAR T-cell Activity

Vascularized microfluidic chip to investigate CAR T-cell migration and cytotoxicity. A, Design of the tri-channel microfluidic device with a 2 mm well in the central channel. B, Schematic diagram showing the development of ovarian cancer-on-a-chip model. Scale bar, 600 μm. C, Immunofluorescence image showing microvasculature in fibrin and OvCAR3 collagen gels (n = 3). Red, HUVEC. Scale bar, 100 μm. D, Real-time images showing the luminal flow of CAR T cells (arrows) through the vasculature formed within the microfluidic device (n = 2). Green, CAR T cells. Scale bar, 20 μm. E, Immunofluorescence image showing an ovarian cancer-on-a-chip 3 days after CAR T-cell treatment. CAR T cells (arrows) migrated into the OvCAR3 gel in the middle of the device. Dotted line marks the edge of the central well. Green, CAR T cells; red, HUVEC. Scale bar, 100 μm. F and G, CD3 (F) and caspase‐3 (Casp3; G) staining and quantification of OvCAR3 gels isolated from microfluidic device after CAR T-cell and anti-TNFα treatment. H, MSD data showing TNFα, IFNγ, and IL2 concentrations on media from microfluidic devices after CAR T-cell and anti-TNFα treatment. F–H, Data plotted as mean ± SD of two/three gels per two replicates. Two different CAR T-cell donors were used in this experiment. Scale bar, 50 μm. Statistics performed using two-way ANOVA.