K562 erythroleukemia cells

MDA-MB-453, MDA-MB-468 and JIMT-1 breast carcinoma, LN-229 glioblastoma, SK-OV-3 ovarian carcinoma, HEK 293 T embryonic kidney (all ATCC, Manassas, VA) and MZ-Mel-2 melanoma cells (kindly provided by Elke Jäger, Krankenhaus Nordwest, Frankfurt, Germany) were cultured in DMEM medium (Gibco, Thermo Fisher Scientific, Darmstadt, Germany), SK-BR-3 breast carcinoma cells (ATCC) in Advanced DMEM/F12 medium (Gibco, Thermo Fisher Scientific), and K562 erythroleukemia cells (ATCC) in RPMI 1640 medium (Gibco, Thermo Fisher Scientific). All media were supplemented with 10% heat-inactivated FBS (Capricorn Scientific, Ebsdorfergrund, Germany), 2 mM L-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin (all Gibco, Thermo Fisher Scientific). Expi293F cells were cultured in Expi293 expression medium (both Gibco, Thermo Fisher Scientific). NK-92 cells [20 (link)] (kindly provided by NantKwest, Inc., Culver City, CA) and ErbB2-specific NK-92/5.28.z cells [21 (link), 22 (link)] were cultured in X-VIVO 10 medium (Lonza, Cologne, Germany) supplemented with 5% heat-inactivated human AB plasma (German Red Cross Blood Donation Service Baden-Württemberg-Hessen, Frankfurt, Germany) and 100 IU/mL IL-2 (Proleukin; Novartis Pharma, Nürnberg, Germany).

EL4 T cell lymphoma, HEK293T embryonic kidney, MDA-MB453 breast carcinoma (all ATCC, Manassas, VA), and SK-MEL-23 melanoma cells [33 (link)] were propagated in DMEM medium (Gibco, Thermo Fisher Scientific, Darmstadt, Germany), UKF-NB3 neuroblastoma cells [34 (link)] in IMDM medium (Gibco) and K562 erythroleukemia cells (ATCC) in RPMI 1640 medium (Gibco). All cell culture media contained as supplements 10% heat-inactivated FBS (Capricorn Scientific, Ebsdorfergrund, Germany), 2 mM L-glutamine, as well as 100 U/mL penicillin and 100 µg/mL streptomycin (all Gibco). NK-92 cells [35 (link)] (kindly provided by NantKwest, Inc., Culver City, CA, USA) were grown in X-VIVO 10 medium (Lonza, Cologne, Germany) with 5% heat-inactivated human AB plasma (German Red Cross Blood Donation Service Baden-Württemberg-Hessen, Frankfurt, Germany) and 100 IU/mL IL-2 (Proleukin; Novartis Pharma, Nürnberg, Germany) as supplements. Peripheral blood mononuclear cells (PBMCs) of healthy donors were isolated from buffy coats using a Ficoll–Hypaque density gradient centrifugation. Peripheral blood NK cells (pNK) were derived from buffy coats using the RosetteSep human NK cell enrichment cocktail (STEMCELL Technologies, Cologne, Germany) following the manufacturer’s recommendations. PBMCs and pNK cells were cultivated in X-VIVO 10 medium with 5% heat-inactivated human AB plasma as a supplement.