NK Cell-Mediated Cytotoxicity Assay

hNK, KHYG-1 and NK-92 cells were co-cultured with NSCLCs: NCI-H1299 or NCI- H1975 at 2.5:1 effector (NK) to target (NSCLC) ratio as was previously determined by the group (Verma et al., 2020 (link)). hNK or NK-92 were cocultured with breast cancer cell lines MCF7 or MDA-MB-231 at 2.5:1 effector (NK) to target (breast cancer cell) ratio. Coculture was carried out in NK MACS media supplemented with 25 ng/ml IL2 for primary human NK cells, or 10% RPMI supplemented with 10 ng/ml IL-2 for KHYG-1 and NK-92 cell lines. hNK, KHYG-1 and NK-92 cell lines were pre-stained with CFSE (Sigma), which was able to perpetuate for 6 days as determined by CFSE + population from flow cytometry, following the manufacturer’s protocol for all co-culture setups except for those used for cytotoxicity assay against model target cell line, K562.