Opto™ Cell Therapy Development Workflow: Cytotoxicity Assay

T cell-mediated tumor death relies on complicated cell-cell interactions and multiple complex mechanisms including the perforin/granzyme pathway, cytokine-dependent killing, and interactions between Fas (T cell) and Fas ligand (tumor cell). To understand and define  the heterogeneity of such interactions, single T cells are penned and cocultured with fluorescently labeled target cells. Killing activity is indicated by caspase-3 activation in target cells. Time-lapse imaging is used to assess killing kinetic. Individual T cells of interest can then be exported alive for downstream analysis such as sequencing. This assay can be combined with other assays in the Opto™ Cell Therapy Development workflow to directly link phenotype and function to gene expression.