Hcc1500

293T (CRL-3216), MCF10A (CRL-10317), T47D (HTB-133), MCF7 (HTB-22), MDAMB231 (CRM-HTB-26), MDAMB468 (HTB-132), BT549 (HTB-122), HCC1500 (CRL-2329), HCC1569 (CRL-2330) and HCC1806 (CRL-2335) were purchased from American Type Culture Collection (ATCC). HCC70 and MDAMB175 were provided by Wake Forest Institutional Cell Bank Repository. Human pre-adipocytes BR-F was obtained from Zen-Bio (BR-F). Human preadipocytes cell line SGBS was a kind gift from Dr. Martin Wabitsch (Ulm University Medical Center). MCF10A was cultured as described in human mammary epithelial growth medium (Lonza)^{78 (link)}. 293T and breast cancer cell lines MCF-7, were cultured in DMEM (Gibco) with 10% fetal bovine serum (FBS). MDAMB468 and MDAMB175-VII was cultured in L15 medium (Gibco) with 10% FBS. MDAMB231, BT549, HCC70, HCC1500, HCC1569, HCC1806, T47D, were cultured in RPMI 1640 (Gibco) with 10% FBS. BR-F was cultured in Preadipocyte Medium (PM-1, Zen-Bio). SGBS was cultured in 0F (DMEM/F12 + pantothenate +biotin + penicillin/streptomycin) plus 10% FBS. All cell lines were authenticated and were regularly tested for mycoplasma contamination.

MCF7, SKBR3, HCC1500, ZR-75-1 (all EpCAM^pos) and MDA-MB-231 (EpCAM^low/neg) breast cancer cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, US). TMX2-28 cells (EpCAM^pos) were a generous gift from Prof. K.F. Arcaro (University of Massachusetts, MA, US) [31 (link)]. All cell lines were cultured in RPMI 1640 containing 10% (v/v) fetal calf serum and 1% (v/v) penicillin/streptomycin (all Gibco/Life Technologies, Darmstadt, Germany). SKBR3 and ZR-75-1 cells were cultured without any supplements, whereas culture media for MCF7 and TMX2-28 were supplemented with 25 mM HEPES. MDA-MB-231 cells were maintained with supplementation of 2 mM L-glutamine and 20 mM HEPES; for HCC1500 cells 2 mM L-glutamine, 1 mM sodium pyruvate (all Gibco/Life Technologies) and 0.45% (v/v) D-(+) glucose solution (Sigma-Aldrich, Munich, Germany) were added. All cells were cultured at 37°C in a humidified atmosphere with 5% CO₂. The same culture conditions were fulfilled for cells grown on coverslips in order to check for marker expression by immunofluorescence staining. At approx. 80% confluence, cells were washed once with PBS (Life Technologies) and fixed with ice cold methanol. Until further use, coverslips were stored at -20°C.

Human BC cell lines HCC‐2218, T47D, BT474, MCF‐7, CAMA‐1 and the immortalized human breast cell line MCF 10A were obtained from the American Type Culture Collection (ATCC). Immortalized HuMEC cells, MDA‐MB‐231 and MDA‐MB‐468 were provided by (Qatar University, Qatar), while HCC‐1500 cell line was obtained from (Weill Cornell Medical College, Qatar). HCC‐2218, T47D, BT474, MCF‐7 and CAMA‐1 cell lines were grown in DMEM medium (Gibco), while MDA‐MB‐231, MDA‐MB‐468 and HCC‐1500 cells were cultured in RPMI 1640 medium. Culture media were supplemented with 10% heat‐inactivated Foetal Bovine Serum (FBS) (Thermo Scientific) and 1% of an antibiotic suspension (Penicillin and streptomycin, Gibco). MCF 10A non‐pathogenic breast cell line was grown in HuMEC Basal serum‐free medium (Gibco, 12753018) supplemented with HuMEC Supplement Kit (Gibco, 12755013), while HuMEC cells were cultivated in keratinocyte serum‐free medium (SFM) supplemented with bovine pituitary extract (BPE) (Gibco, 17005042). All cells were maintained with an atmosphere of 5% CO₂ in a humidified incubator adjusted to 37°C.