Cytotoxic T cell based Tumor Killing Assays

Charles River’s tumor killing assays include:

  • IncuCyte tumor killing assay (2D)
  • Macrophage-dependent ADCC
  • IncuCyte spheroid tumor killing assay (3D)/CDC
  • Flow cytometry-based killing assays

IncuCyte based tumor killing assay enables quantification of target number that can be multiplexed with target specific apoptotic readout. This assay can be tailored to the anticipated mechanism of action by selecting from a panel of validated target cell lines.

Cytotoxic T Cells can be cultured with activating antibodies such as anti-CD3 and tested for their ability to kill tumor targets in an IncuCyte based assay. The ability of novel therapeutics to potentiate cytotoxic T cell killing can then be assessed by quantifying the change in the number of viable tumor cells over time. Positive control agents which enhance tumor killing have been validated in this assay. Caspase 3/7-dependent tumour cell apoptosis can also be determined by co-localisation of the caspase signal to NucLight Red positive tumor cells. The target cell population can be adapted to suit the disease indication or mechanism of action.

PBMC - mediated tumor killing in 2D format, and example image, demonstrating killing over time with Pembrolizumab/Ipilimumab with or without depletion of CD8+ cytotoxic T cells.
Figure 1: PBMC - mediated tumor killing in 2D format, and example image, demonstrating killing over time with Pembrolizumab/Ipilimumab with or without depletion of CD8+ cytotoxic T cells.

Charles River also run immune cell mediated tumor killing assays with PDX (patient derived xenograft) material; in vitro 3D assays incorporate live cell imaging to predict in vivo responses to matched PDX material. Screening PDX models in vitro prior to in vivo humanized tumor killing models enables stratification and prioritization of the wide range of in vivo PDX models available. The combination of in vitro screening with in vivo trial design options will aid your drug development and increase throughput.