title: Single-colour flow cytometric assay to determine NK cell-mediated cytotoxicity and viability against non-adherent human tumor cells.
abstract: A flow cytometry-based cytotoxicity (FCC) assay was developed using a single fluorophore, calcein-acetoxymethyl diacetylester (calcein-AM), to measure NK cell-mediated cytotoxicity. Non-adherent human K562 and U937 target cells were individually labelled with calcein-AM and co-incubated with effector NK cells to measure calcein loss, and therefore calculate target cell cytotoxicity. This FCC assay also provided a measure of sample viability. Notably, cell viability measured by traditional calcein/7-amino-actinomycin D (7-AAD) double labelling and Trypan Blue methods were comparable to the viability calculated using calcein-loss FCC. This FCC assay may also be used with various effector and target cell types and as a multi-parameter tool to measure viability and immunophenotype cells for tissue engineering purposes.
authors.affiliation: School of Biomedical Engineering, McMaster University, 5380 Glen Erin Dr., Mississauga, ON L5M5C7, Canada. thakurajit@gmail.com
authors.forename: Ajit
authors.initials: A
authors.lastname: Thakur
authors.forename: Abeyat
authors.initials: A
authors.lastname: Zaman
authors.forename: Jeff
authors.initials: J
authors.lastname: Hummel
authors.forename: Kim
authors.initials: K
authors.lastname: Jones
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Hortelano
mesh_terms: D002470:Cell Survival; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D013194:Staining and Labeling / Q000379:methods; D014407:Tumor Cells, Cultured
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; C085925:calcein AM