Hek 293 t embryonic kidney

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).

Human PC-3 prostate cancer (Cat# CRL-1435), Jurkat T lymphoma (TIB-152), and HEK-293T embryonic kidney (CRL-3216) cells were purchased from ATCC, while donated primary CD3⁺ samples of T cells from three different donors were purchased from Cellero (formerly known as Astarte Bio). PC-3, Jurkat, and HEK-293T cells were cultured in serum-containing RPMI-1640 media that was supplemented with 10% fetal bovine serum, except during the 24-h period after a transfection (unless otherwise noted). In contrast, primary T cells were cultured in serum-free X-VIVO15 media that was supplemented with 0.5 ng/mL IL-2 and anti-CD3/28 Dynabeads in a 1:1 cell:bead ratio. Dynabeads were replaced weekly, while IL-2 was added to the media every 2–3 days.