Ecacc85090402

The human epidermis carcinoma cell line A431 (ECACC85090402) was obtained from the European Collection of Authenticated Cell Cultures through DS Pharma Biomedical (Osaka, Japan). The mouse embryo fibroblast cell line BALB/3T3 clone A31 (JCRB9005) was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). The human breast ductal carcinoma cell line BT-474 (HTB-20) was obtained from the American Type Culture Collection through Summit Pharmaceuticals International Corporation (Tokyo, Japan). These cell lines were cultured in Dulbecco’s modified Eagle’s medium (10569010; Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (26140079; Thermo Fisher Scientific) and 1% penicillin/streptomycin (15140122; Thermo Fisher Scientific). Each cell line was cultured in tissue culture flasks at 37 °C in a 5% CO₂ humidified atmosphere.

A series of human cancer cell lines (A549, A431, PC3, MCF-7, SK-BR-3, MDA-MB-23, HT-29 and HCC-2998) were screened regarding the expression of CCK-2R. The A549 non-small cell lung cancer and A431 skin cancer cell lines were purchased from the European Collection of Authenticated Cell Cultures (ECACC 86012804 and ECACC 85090402). The PC3 prostate cancer cell line; the MCF-7, SK-BR-3, and MDA-MB-231 breast cancer cell lines; and the HT-29 and HCC-2998 colon cancer cell lines were kindly provided by Dr. Alessia Ciarrocchi (Laboratory of Translational Research AUSL-IRCCS, Reggio Emilia, Italy). A549, A431, PC3, MCF-7, and HCC-2998 were cultured in RPMI-1640 added of 10% heat-inactivated FBS. SK-BR-3, MDA-MB-231, and HT-29 were cultured in DMEM with high glucose added to 10% heat-inactivated FBS. Media contained 100 U/mL penicillin-streptomycin and glutaMAX. Cell culture reagents were purchased from Thermo Fisher (Milan, Italy). Cell lines were routinely maintained at 37 °C with 5% CO₂.