Ecb9006l

Hep3B and HepG2 cell lines were purchased from the Leibniz-Institut DSMZ, Braunschweig (ACC-93, ACC-180) and grown in DMEM (Euroclone, ECM0749L) and RPM1640 (Euroclone, ECB9006L), respectively, supplemented with 10% FBS (Euroclone, ECS0180L) and 1% penicillin-streptomycin. HuS cells were a gift of Dr. Vinicio Carloni, University of Florence and were grown in DMEM containing 4.5 g/L of glucose (Euroclone, ECB7501L) and supplemented with 10% FBS (Euroclone, ECS0180L), 5 ng/mL EGF (Sigma-Aldrich, E9644), 420 ng/mL insulin (Sigma-Aldrich, I9278), 20 ng/mL selenium (Sigma-Aldrich, S9133), 1% DMSO (Amresco, 0231) and 1% penicillin-streptomycin (Euroclone, ECB3001D). Cells have been authenticated by STR PCR by the supplier and routinely tested for Mycoplasma contamination using Myco Alert (Lonza, LT07-318).

Human osteosarcoma cells U2OS, human embryonic kidney cells 293 (HEK293), human melanoma cancer cells A375 and human urinary bladder epithelial carcinoma cells (5637, HTB9) were acquired from American Type Culture Collection and the National Collection of Type Cultures (ATCC), were grown in Dulbecco’s modified Eagle’s medium (DMEM, Euroclone, ECB7501L) with 4.5 g/L glucose supplemented with 10% fetal bovine serum (FBS, Gibco, 10270106), 2 mM L-glutamine and 1% Penicillin/streptomycin. Breast cancer cells, MDA-MB-231, were cultured in DMEM with 4.5 g/L glucose supplemented with 5% FBS, 4mM L-glutamine and 1% Penicillin/streptomycin. Bladder carcinoma cells, 5637, and human keratinocyte NCTC2544 were cultured in RPMI 1640 medium (Euroclone, ECB9006L) supplemented with 10% FBS and 2 mM L-glutamine. All cell lines were grown at 37 °C in a humidified atmosphere with 5% CO₂.