Hcc1599

HCC1806, MDA-MB-436, HCC1395, ZR-75-1, T47D, MCF7, BT474, and MCF10A were a kind gift from CRUK Manchester Institute and they were authenticated by short tandem repeat profiling by CRUK MI core facility unit. HCC1806 was authenticated again in August 2017 by ATCC Service. HCC1569 and HCC1599 were purchased from ATCC in April 2016. MDA-MB-453 and CAL-85-1 were purchased from DSMZ in November 2014 and November 2016, respectively. MDA-MB-321 and BT20 was a kind gift from Department of Immunology, Medical University of Warsaw (purchased from ATCC in February 2018). ATCC and DSMZ authenticate cell lines by short tandem repeat profiling. HCC1806, HCC1395, BT474, BT20, ZR-75-1, T47D, HCC1599 and HCC1569 were cultured in RPMI-1640 supplemented with 10% FCS, 1% penicillin/streptomycin, 2 mM l-glutamine and 1 mM sodium pyruvate. MCF7, CAL-85-1, MDA-MB-453 and MDA-MB-231 were cultured in DMEM supplemented with 10% FCS, 1% penicillin/streptomycin, 2 mM l-glutamine and 1 mM sodium pyruvate. MDA-MB-436 were cultured in RPMI-1640 with 25 mM HEPES supplemented with 10% FCS, 1% penicillin/streptomycin, 2 mM l-glutamine, 1 mM sodium pyruvate and 10 µg/ml insulin. MCF10A were grown in DMEM/F12 supplemented with 5% horse serum, 20 ng/ml EGF, 0.5 mg/ml hydrocortisone, 100 ng/ml cholera toxin, 10 µg/ml insulin, 1% penicillin/streptomycin.

MCF-10A, MCF-7, HCC1599, MDA-MB-231 and SK-BR-3 were obtained from ATCC (American Type Culture Collection) and SUM159PT was supplied by Aster-and Bioscience. The cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and maintained in a CO2 incubator at 37°C. The MycoSEQ™ Mycoplasma Detection Kit (#4460623, Thermo Fisher Scientific) was utilized to detect mycoplasma contamination in the cell culture. Before we began our experiments, a short tandem repeat (STR) profile test was used to formally qualify the cell lines.

PC-3, 22Rv1, LNCaP, VCaP, DU145, MDA-MB-231, ZR-75-1, BxPC3, 786-O, A498, HCT-116, HT-29, A2058, SkMel2, A375, OV90, PANC1, HCC1187, TCCSUP, RT4, HepG2, DU4475, HCC1806 and HCC1599 were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cell lines derived from PC-3 (PC-3E and TEM 4-18^{9 (link)}; GS689Li and GS694Lad^{9 (link)}; TEM 4-18 ZEB1 KO) were previously generated in our laboratory. The identity of PC-3E, TEM 4-18, GS689Li and GS689Lad as PC-3 derivatives wes confirmed by STR profiling (IDEX Laboratories). All cell lines were grown at 37 °C in a 5% CO₂ atmosphere in ATCC-specified medium containing 10% fetal bovine serum and 1% non-essential amino acids. Monensin (M5273, 90–95% purity), salinomycin (S4526), nigericin (N7143), pyrvinium pamoate (1592001), epirubicin hydrochloride (E9406), gentian violet (48770), dactinomycin (1162400) and daunorubicin (D8809) were purchased from Sigma (Saint-Louis, MO).

The breast cancer cell lines (BT-20, HCC70, HCC1187, HCC1599, MDA-MB-468 and MDA-MB-436) used for establishing the Notch transcriptomic signature were purchased fresh from the American Type Culture Collection (ATCC) and maintained in RPMI 1640 medium (ATCC’s modification, cat #A1049101, Thermo Fisher Scientific) with 10% fetal bovine serum (FBS, heat inactivated, cat #A3840402, Thermo Fisher Scientific) and 1% penicillin–streptomycin (cat #15140-122, Thermo Fisher Scientific). The 19 breast cancer cell lines used to select the final signature were maintained as described [46 (link)]. Accutase (cat # A6964-100ML, Sigma-Aldrich) was used to detach the cells prior to seeding on coated plates and during maintenance. All cell lines were cultivated with 5% CO2 at 37 °C.