Sk br 3

Human breast cancer cell lines T-47D, MCF7, MDA-MB-361, SK-BR-3, HCC1954, MDA-MB-468, MDA-MB-231, and BT-549 were obtained from the ATCC (Manassas, VA, USA). MCF7, MDA-MB-361, SK-BR-3, and MDA-MB-231 cells were cultured in Dulbecco modified Eagle medium (Lonza, Basel, Switzerland), and T-47D, HCC1954, MDA-MB-468, and BT-549 were maintained in Roswell Park Memorial Institute-1640 medium (Lonza); all media were supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA), penicillin (100 units/mL), and streptomycin (100 mg/mL). All cells were cultured at 37 °C in a humidified atmosphere of 5% carbon dioxide.

NCI-H1993 (CRL-5909), NCI-H441 (HTB-174), SKBR3 (HTB-30), MKN-45 (ACC409), OE33 (ACC706), MCF7 (HTB-22), MDA-MB-231 (HTB-26) and NIH3T3 (CRL-1658) cell lines were obtained from ATCC (http://www.lgcstandards-atcc.org/en.aspx) or DSMZ (http://www.dsmz.de/), tested for mycoplasma infection on a regular basis using a commercial biochemical test (Lonza) and authenticated using STR profiling. All cells were cultured in Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham, 1:1 mixture supplemented with 10% fetal calf serum (FCS), or in case of MKN45 with 20% FCS. Upon serum withdrawal, FCS was replaced by 0.1% bovine serum albumin.

The breast cancer cell lines MCF7, T47D, MDA-MB-231, SK-BR-3, MDA-MB-453, MDA-MB-468, Hs578T and MCF10A were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA), where they are regularly authenticated. The cell lines were grown at 37°C in 5% CO₂. Hs578T, MDA-MB-231, MDA-MB-453, MDA-MB-468, and SK-BR-3 cells were maintained in DMEM (Lonza, Basel, Switzerland) containing 10% fetal bovine serum (FBS, Gibco BRL, Grand Island, NY, USA), penicillin (100 units/ml; Gibco) and streptomycin (100 units/ml, Gibco). MCF7 and T47D cells were cultured in RPMI 1640 (Lonza), 10% FBS, penicillin (100 units/ml) and streptomycin (100 units/ml). MCF10A cells were cultured in DMEM/F12 media (1:1) (Invitrogen, Grand Island, NY, USA) supplemented with 5% horse serum (Gibco), 10 μg/ml bovine insulin (Sigma-Aldrich, St. Louis, MO, USA), 20 ng/ml epidermal growth factor (EGF; Sigma), 0.5 μg/ml hydrocortisone (Sigma), 0.1 μg/ml cholera toxin (Sigma), penicillin (100 units/ml), and streptomycin (100 units/ml). Human umbilical vein endothelial cells (HUVECs; Lonza) were maintained in Lonza EGM-MV (normal growth medium) at 37°C in 5% CO₂. The cells were maintained in culture plates and used in assays between cell passages 3 and 8.

Human breast cancer cells MDAMB231, MDAMB468, MCF-7, SKBR3 and MDAMB361 were purchased from American Type Culture Collection (Rockville, MD, USA). All cell lines were genotyped to confirm their origin. MDAMB231, MCF7 and SKBR3 cells were maintained in DMEM high glucose (Lonza) complemented with 10% fetal bovine serum (FBS; Lonza). MDAMB468 and MDAMB361 were maintained in DMEM-F12 complemented with 10% FBS. All media were supplemented with 10000 U/ml penicillin and 10 mg/ml streptomycin (Lonza) and 4 mM L-glutamine in a humidified atmosphere composed of 95% air and 5% CO2 at 37  °C. Cell lines were regularly inspected for mycoplasma.

Human breast cancer cell lines MDA-MB-231, MCF-7, BT474, SK-BR-3, ZR-75–1 together with normal breast epithelial cell lines MCF-10A and MCF-12A were obtained from ATCC (Manassas, VA, USA). MDA-MB-231 and SK-BR-3 cell lines were cultured with Dulbecco Modified Eagle Medium (Lonza, NJ, USA) while MCF-7, BT474 and ZR-75–1 cell lines were cultured with DMEM supplemented with 0.1% insulin. MCF10A and MCF-12A cell line was cultured with DMEM (Lonza, NJ, USA) supplemented with 0.1% insulin (0.01 mg/ml), 0.002% EGF (20 ng/ml) (both from Sigma-Aldrich, Saint-Louis, MO, USA). All media were supplemented with 50 U/ml penicillin/streptomycin, 1% non-essential amino acids and 10% fetal bovine serum (Lonza, NJ, USA). All cell lines were tested for mycoplasma contamination regularly using MycoAlert mycoplasma detection kit (Lonza, NJ, USA).

The prostate carcinoma cell lines PC3 and LNCaP and breast carcinoma cell lines MCF7 and SKBR3 were obtained from ATCC (Manassa, VA, USA). The PC3-9 cell line^{24 (link)}, a sub-clone of the PC3 cell line, was kindly provided by Immunicon (Huntingdon Valley, PA, USA). PC3, PC3-9 and LNCaP were cultured in RPMI1640 (Lonza), MCF7 and SKBR3 in DMEM (Lonza), all supplemented with 10% FBS (Sigma) and 1% Pen/Strep (Lonza). Cells were cultured at 37° in a humid atmosphere and trypsinized when reaching 70–80% confluence using 0.05% trypsin–EDTA (Gibco, Life Technologies, Waltham, MA, USA).

Human breast cell lines (BCCLs), SkBr3, MDA-MB-468, T47D, purchased from the American Type Culture Collection, were cultured in McCoy’s 5A (SkBr3) or Dulbecco’s modified Eagle’s medium (MDA-MB-468, T47D) (Lonza, Slough, UK) supplemented with 5% (T47D) or 10% fetal bovine serum (Lonza). The human normal fibroblast (NF) cell line NHDF (normal human dermal fibroblasts), derived from human normal derma (Lonza) was cultured in Fibroblast Basal Medium (FBM) supplemented with Fibroblast Growth Medium-2 (FGM-2) Bullet kit (Lonza), containing 2% fetal bovine serum (FBS), 0.1% Insulin, 0.1% gentamicin, amphotericin GA 1000, 0.1% fibroblast growth factor (FGF). All cell lines were cultured at 37 °C in 95% humidified air in the presence of 5% CO₂ and authenticated with short tandem repeat DNA profiling analysis by the Functional Genomic Unit of the Department of Experimental Oncology at Fondazione IRCCS Istituto Nazionale Tumori of Milano (INT).