Cytotoxicity is an essential parameter to examine killer cell activity, to screen hits for new drugs, and to proof safety evaluation. Cytotoxicity assays are routinely used in basic research, clinical studies, and industrial applications. However, the most commonly applied standard end-point cytotoxicity assays largely suffer from the lack of kinetic information, which is indispensable to assess killing competence. Here we report a quantitative and highly sensitive calcein-based real-time high-throughput cytotoxicity assay on a single cell level and for population use. By determining fluorescence loss as a function of the fraction of lysed cells, this assay measures the kinetics of cytotoxicity over a broad dynamic range. The high time resolution of the assay also provides kinetic information with the potential to reveal neglected/immeasurable properties in the process of cytotoxicity of both purified human cytotoxic T lymphocytes and natural killer cells. To test its clinical potential, we have also applied this assay to screen the cytotoxic potential of (easy to prepare) peripheral blood mononuclear cells in healthy blood donors. In summary, this assay should substantially increase the screening efficiency for clinical and industrial purposes and offer quantitative kinetic data with a high time resolution.
- Copyright © 2013 by The American Association of Immunologists, Inc.