Mco 17ai

The human breast cancer cell line (MCF-7) and normal fibroblast mouse (L929) cell (as control) were obtained from the Pasteur Institute of Iran and cultured in RPMI medium supplemented with 10% FBS, 100 U/ml penicillin, and 100 mg/ml streptomycin. The MCF-7 cells were cultured in a CO₂ incubator (MCO-17AI, Sanyo Electric Co., Ltd., Japan) at 37°C in a humidified atmosphere enriched by 5% CO₂ and subcultured every 3–4 days. The malignant and nonmalignant cells were cultured in Dulbecco's modified Eagle's medium with 5% (v/v) FBS, 100 units/ml penicillin and 100 μg/ml streptomycin.

Experiments were carried out using mouse neuroblastoma N2A cell line (tumor-like neuroblasts), which were kindly provided by Dr. M. Mojarad. The cells were cultured either in 96-well tissue (TC) plate (NUNC, Wiesbaden, Germany) or in 25-cm² TC flasks (NUNC, Wiesbaden, Germany). The cells were cultured in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% fetal calf serum (Gibco–Invitrogen), 100 IU·ml^-1 penicillin (Gibco–Invitrogen), and 100 μg·ml^-1 streptomycin (Gibco–Invitrogen). The cells were incubated in a humidified 5% CO₂ and 95% air atmosphere at 37°C in CO₂ incubator MCO-17AI (Sanyo Electric Co., Ltd., Japan). The culture medium was replaced 3 times a week. At the time of the experiment, confluent cells were trypsinized and plated in 96-well plates or into tissue culture dishes (40 mm in diameter; TPP). N2A cells were plated at a density of 31, 000 cells/cm². Experiments were initiated 72 h after plating.