Cell culture

Prostate adenocarcinoma cell lines PC3 (ATCC #CRL-1435), DU145 (ATCC #HTB-81) and prostate carcinoma cell line LNCaP (ATCC #CRL-1740), were purchased from American Type Culture Collection (Manassas, VA, USA). PC3, DU145 and LNCaP cells were cultured in RPMI 1640 cell culture medium (Invitrogen, Grand Island, NY, USA). Medium was supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) PenStrep, all purchased from Invitrogen. PC3, DU145 and LNCaP cells were incubated under humidified conditions at 37°C and 5% CO₂. For passaging, the cancer cells were lifted by washing the flasks with HBSS buffer without calcium and magnesium (Invitrogen). After washing, the cells were incubated with trypsin and 0.05% EDTA (Invitrogen) for 2–5 min. Complete RPMI 1640 was added to the cells and they were collected into a 15 ml tube. The cells were then centrifuged at 300 g for 5 min. The supernatant was removed, and the cells were reseeded into flasks or used for experiments.