Simultaneous measurement of target and effector cell death, apoptosis, and proliferation in cell therapy development using flow cytometry

The development of cellular immunotherapies against cancer requires an efficient assessment of malignant target cell killing. However, the fate of effector cells is equally important, particularly in the context of immunomodulatory tumors. Here, we present a high-throughput protocol for simultaneously measuring viability, apoptosis, and proliferation of cytotoxic effector cells and their targets. We describe steps for co-culturing target and effector cells, staining procedures for viability and apoptosis, adding fluorescent counting beads for cell quantification, and procedures for flow-cytometric analysis.