• Functional endpoints: T cell infiltration and tumor killing

Two breast cancer cell lines (ZR-75-1 and SKBR-3) were grown in 3D to form tumoroids. HLA-matched PBMCs from a healthy donor were added. Cells were co-cultured in the absence or in the presence of T cell stimulator anti-CD3/28 beads or superantigen SEB. Invasive ratio was used as a measure of T cell infiltration. Blue bars represent tumoroid volumes in monocultures (without PBMCs). Remaining total tumoroid volume was used as a read-out of T cell mediated killing.

 Physiologically relevant environment

3D environment allows different cell types to engage in a more realistic spatial context than when cultured on a plastic dishes. Subsequent steps that occur during tumoroid-T cells encounter can be dissected using 3D high content microscope.

 

Multiple donors to address donor-to-donor variability

PBMCs from different donors of the same HLA-type were co-cultured with SKBR-3 breast cancer cell line grown in 3D and their infiltration and killing capacity was measured after stimulation with superantigen SEA.

Control the strength of the anti-tumor immune response

Different target-to-effector ratios and different activation stages of immune cells can be tested. Bi-parametric analysis of tumoroids size vs T cells invasiveness allows to better understand drugs immuno-modulatory profile: do they impact on infiltration/killing or both? 

Flexible design of co-cultures

Assays can be easily customized  by replacing the cellular players. This allows for testing whole range of immunotherapies that focus on different immune compartments and target diverse cancer indications.