Quantifying T Cell Response in 3D Tumor Spheroids Using Advanced Flow Cytometry Workflows
This application note provides two Advanced Flow Cytometry Platform-based solutions to address problems with traditional methods for measuring T cell response in 3D models.
A tumor infiltrating lymphocytes (TILs) assay to measure the degree of T cell infiltration into spheroids, and to determine the phenotype of infiltrated compared to non-infiltrated cells, is described.
Also detailed is the spheroid immune cell killing (ICK) workflow, which uses subset analysis and cytokine quantification, to examine the activation, killing, exhaustion and memory profiles of T cells during co-culture with spheroids.
Both solutions utilize the iQue® Advanced Flow Cytometry Platform validated suite of T cell characterization reagent kits and validated spheroid washing and dissociation protocols, to provide an end-to-end solution for the evaluation of T cell response in advanced 3D tumor models.
In addition to assay principles, this application note includes the following sections:
- Activation marker expression on TILs is greater than on non-infiltrated T cells
- Tumor cell type impacts the number and profile of TILs
- Activation and exhaustion status of T cells changes throughout spheroid killing
- Stimulated T cells shift from early to late stage memory cell development
- Anti-HER2 CAR-T killing is specific to HER2 Positive tumor spheroids