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-Saenz

doi:10.1016/j.xpro.2023.102224

A co-culture model system to quantify antibody-dependent cellular cytotoxicity in human breast cancer cells using an engineered natural killer cell line

STAR Protoc. 2023 Apr 17;4(2):102224. doi: 10.1016/j.xpro.2023.102224. Online ahead of print.

Authors

Roos Vincken¹, Ana Ruiz-Saenz²

Affiliations

¹ Department of Cell Biology, Erasmus University Medical Center Rotterdam, 3015 CN Rotterdam, the Netherlands. Electronic address: r.vincken@erasmusmc.nl.
² Department of Cell Biology, Erasmus University Medical Center Rotterdam, 3015 CN Rotterdam, the Netherlands; Department of Medical Oncology, Erasmus University Medical Center Rotterdam, 3015 GD Rotterdam, the Netherlands. Electronic address: a.ruizsaenz@erasmusmc.nl.

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.

Keywords: Antibody; Biotechnology and bioengineering; Cancer; Cell Biology; Cell culture; Cell isolation; Cell-based Assays; Flow Cytometry/Mass Cytometry; Immunology.