A co-culture model system to quantify antibody-dependent cellular cytotoxicity in human breast cancer cells using an engineered natural killer cell line
Roos Vincken, Ana Ruiz-Sáenz
Abstract
Current protocols measure antibody-dependent cellular cytotoxicity (ADCC) in vitro using peripheral blood mononuclear cells (PBMCs), but isolation and variability among donors limit the viability and reproducibility of this approach. Here, we present a standardized co-culture model system to quantify ADCC on human breast cancer cells. We describe steps to engineer a natural killer cell line that stably expresses FCγRIIIa (CD16), required to mediate ADCC. We then detail the steps for the cancer-immune co-culture setup, followed by cytotoxicity measurement and analysis.
Topics & Concepts
Antibody-dependent cell-mediated cytotoxicityCytotoxicityCD16Peripheral blood mononuclear cellImmune systemCell cultureImmunologyJurkat cellsCancerAntibodyCancer researchNatural killer cellFlow cytometryBiologyCancer cellIn vitroT cellCD3BiochemistryGeneticsCD8CAR-T cell therapy researchMonoclonal and Polyclonal Antibodies ResearchImmune Cell Function and Interaction