Egf af 100 15 1mg

Human glioblastoma cells -U87(ATCC® HTB14™) were obtained from ATCC, MCF10CA1h cells [35] were received from the Barbara Ann Karmanos Cancer Institute (Detroit, MI, USA), and they were cultured as previously described. Briefly, the U87 cells were cultured in DMEM -Dulbecco's Modified Eagle Medium (ThermoFisher Scientific, 11995065) that was supplemented with 10% fetal bovine serum (FBS) (ThermoFisher Scientific, 11995065) and 50 U/mL penicillin and 50 µg/ml streptomycin (Thermo Fisher, 15070-063). MCF10CA1h cells were cultured in the complete medium: DMEM/F12 (11330-032, Thermo Fisher Scientific), 5% horse serum (16050-122, Thermo Fisher Scientific), 5 ng/ml EGF (AF-100-15-1MG, Peprotech), 0.5 mg/ml Hydrocortisone (H0888-1G, Sigma-Aldrich), 100 ng/ml Cholera toxin (C8052-2mg, Sigma-Aldrich), 10 µg/ml insulin (I1882-100MG, Sigma-Aldrich) and 1x penicillin/streptomycin solution (15070-063, Thermo Fisher Scientific). Cells were cultured at 37˚C, in 5% CO2. Cell propagation was performed by detaching adherent cells using Trypsin (0.25% for U87 cells (ThermoFisher Scientific, 80-2101) and 0.05% for MCF10CA1h cells (25-052-Cl, Corning)) as previously described. All experiments were performed using cells with passage number less than 19. Cell medium was changed every 2-3 days.