Method Profile
Measure organoid number and area over time, then combine those data with T-cell proximity and endpoint flow cytometry.
Compare candidate CAR-T activity across patient-derived pancreatic cancer organoids.
Seven PDAC organoids with healthy-donor CAR or nontransduced T cells; CD318 and TSPAN8 target comparisons used relevant subsets.
Pretrained OrganoIDNet organoid segmentation, a separate T-cell detection pipeline and endpoint cytometry.
Tumor models
Patient-derived pancreatic ductal adenocarcinoma organoids in a Matrigel-based coculture.
Immune cells
CAR and nontransduced T cells from three independent healthy donors in the main comparisons.
Specificity controls
Antigen knockout cell lines and TSPAN8-positive/negative organoids.
Interpret organoid morphology, T-cell proximity and endpoint killing as complementary measurements, with technical wells averaged within donor.
Repeated imaging with orthogonal functional and antigen controls.
Two primary PDOs, four additional CD318 models and TSPAN8 organoid comparisons within a seven-model collection.
CAR T cells versus nontransduced T cells or no T cells; antigen-positive versus relevant negative controls.
Main-model killing increased with effector-to-target ratio, reaching approximately 45% and 40% at 8:1 after three days.
Repeated imaging measured changes in organoid number, area and nearby T cells during CAR-T coculture. Flow cytometry and antigen controls provided complementary evidence of tumor-cell killing.
