Mda mb 231

The human breast cancer cell line MDA-MB-231 was purchased from the Korea Cell Line Bank (Seoul, Korea), and RT-R-MDA-MB-231 cells were established as previously described [15 (link)]. In brief, MDA-MB-231 cells were irradiated with X-rays (2 Gy/25 fractions) to a total dose of 50 Gy, a commonly used clinical RT regimen in breast cancer patients. RT-R-MDA-MB-231 cells were used through 5 passages. The human umbilical endothelial cell line EA.hy926 and human NK cell line NK-92MI were obtained from the American Type Tissue Culture Collection (ATCC; Manassas, VA, USA). Breast cancer cell lines and endothelial cell lines were grown in RPMI-1640 medium and DMEM, respectively, supplemented with 10% FBS (GenDEPOT, Katy, TX, USA) and 1% penicillin and streptomycin (HyClone; Danaher Corporation, WA, Washington, DC, USA) at 37 °C in humidified air containing 5% CO₂. NK-92MI cells were grown in alpha-MEM containing 2 mM L-glutamine, 1.5 g/L sodium bicarbonate (#12561-056, GIBCO; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 0.2 mM inositol (#I7508, Sigma-Aldrich, St. Louis, MO, USA), 0.1 mM 2-mercaptoethanol (#M3148, Sigma-Aldrich), 0.02 mM folic acid (#F8758, Sigma-Aldrich), 12.5% horse serum (GIBCO), and 12.5% FBS without ribonucleosides and deoxyribonucleosides at 37 °C in humidified air containing 5% CO₂.

Human gastric cancer cell lines (AGS, SNU638 and SNU719) and breast cancer cell lines (BT549, MCF7 and MDA-MB231), mouse melanoma cell line B16F10 were purchased from the Korean Cell Line Bank (KCLB). Human cancer cell lines were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin/streptomycin (Capricorn scientific) at 37 °C in a humidified atmosphere of 5% CO₂, and B16F10 was cultured in Dulbecco’s Modified Eagle Medium (DMEM) medium under the same conditions. TQ was suspended in dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA).

The human breast cancer cell lines MCF-7 and MDA-MB-231 were obtained from the Korea Cell Line Bank (KCLB, Seoul, Korea). Both cell lines were grown in Dulbecco’s Modified Essential Medium (DMEM; HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; HyClone), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (HyClone), and were maintained at 37 °C in a humidified incubator containing 5% CO₂. We used breast cancer cell lines up to 15 passages after purchasing cell lines from KCLB and the cell lines were authenticated by the KCLB and showed >90% similarity in a short tandem repeats (STR) DNA fingerprint profile when compared with the American Type Culture Collection (ATCC) and Korean Cell Line Bank (KCLB) databases. Cells were plated at a density of 1 × 10⁶ cells in 10-cm culture dishes. To establish primary mammospheres, single-cell suspensions of MCF-7 and MDA-MB-231 cells were seeded at a density of (3.5∼4) × 10⁴ and (0.5∼1) × 10⁴ cells/well, respectively, in ultralow attachment six-well plates containing 2 mL of complete MammoCult^TM medium (StemCell Technologies; Vancouver, BC, Canada), which was supplemented with 4 μg/mL of heparin, 0.48 μg/mL of hydrocortisone, 100 U/mL of penicillin, and 100 μg/mL of streptomycin. The cells were incubated for seven days in a 5% CO₂ incubator at 37 °C.