Mda mb 231 breast carcinoma

MDA-MB-435, A375 melanoma cells and MDA-MB-231 breast carcinoma cell line were purchased from ATCC and maintained in low passage numbers according to ATCC guidelines. All cell lines were maintained in master cell banks and undergo routine mycoplasma testing. Any cells displaying abnormal morphological changes or doubling time are discarded and replaced with a new vial. MDA-MB-435, MDA-MB-231 and A375M cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37°C under 5% CO2. Patient-derived TILs cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% human serum at 37°C under 5% CO2. Human CD8+ T cells were isolated from buffy coats, activated with anti-CD3 and CD28 mAbs that were plate-bound and expanded in RPMI 1640 medium supplemented with 10% human serum and 100 IU/mL IL-2 and 5 ng/ml IL-15 at 37°C under 5% CO2.

A-10 embryonic rat aortic smooth muscle, HeLa cervical carcinoma, SK-OV-3 ovarian carcinoma, and MDA-MB-231 breast carcinoma cell lines were obtained directly from ATCC (Manassas, VA). The MDA-MB-435 melanoma cells were acquired from the Lombardi Cancer Center (Georgetown University) and validated by ATCC. NCI/ADR-RES cells were obtained from the NIH (Bethesda, MD). The SK-OV-3/MDR-1–6/6 and HeLa wild-type βIII (WT βIII) cell lines were described previously.^{17 (link)} MDA-MB-435, MDA-MB-231 and NCI/ADR-RES cells were cultured in Improved Minimum Essential Medium (Richter’s Modification) (Gibco, Life Technologies, Grand Island, NY) with 10% fetal bovine serum and 25 μg/mL gentamicin. WT βIII cells were grown in DMEM (Gibco, Life Technologies) with 10% fetal bovine serum and 50 μg/mL gentamicin, while all the other cell lines were maintained in Basal Medium Eagle (Sigma-Aldrich) with 10% fetal bovine serum and 50 μg/mL gentamicin. All cells were maintained in a humidified incubator at 37 °C with 5% CO₂.