Megm singlequot supplements

Breast cancer cell lines were cultured in RPMI-1640 (Life Technologies, Carlsbad, CA, USA) + 10% heat-inactivated fetal bovine serum (Sigma, St. Louis, MO, USA). Cells were regularly monitored for mycoplasma contamination using MycoAlert kit (Lonza, Walkersville, MD, USA). Cell lines were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA) and authenticated using the STR profiling method by the City of Hope Genomics Core facility. HMECs were cultured in Mammary Epithelial Cell Basal Medium (MEBM) plus addition of Mammary Epithelial Cell Growth Medium (MEGM) SingleQuot supplements (Lonza) to generate complete culture medium.

MDA-MB-231, MCF7, Jurkat E6.1 (European Collection of Authenticated Cell Cultures), HEK293, HL-60, and RAMOS (ATCC) were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), penicillin (50 units/mL), and streptomycin (50 µg/mL) in a humidified incubator at 37 °C with 5% CO₂. MCF10A breast epithelial cell line (ATCC) was cultured in mammary epithelial cell growth medium (MEGM) with MEGM SingleQuot Supplements (hydrocortisone, rhEGF, insulin, and bovine pituitary extract) as instructed (Lonza) and 100 ng/mL cholera toxin (Sigma). Cell lines were passaged by 0.05% trypsin and 0.5 mM EDTA (Gibco) and quenched with serum-containing medium. MCF10A cells were trypsinized and quenched with Defined trypsin Inhibitor (Thermo Fisher Scientific). Cell viability was measured by trypan blue exclusion assay. Briefly, 10 µL of cell suspension was mixed with 10 µL of 4% trypan blue solution and total and trypan-stained cells were quantified by Countess II Automated Cell Counter (Thermo Fisher Scientific).