Ecm0180d

The MeT-5A cell line, derived from pleural fluids obtained from non-cancerous human individuals, was purchased from the European Collection of Cell Cultures (ECACC) and cultured in RPMI 1640 medium (R0883, Sigma) plus 15% fetal bovine serum (ECS0180L, Euroclone, Milan, Italy), 100×U/mL penicillin plus 100 mg/mL streptomycin (ECB3001D, Euroclone, Milan, Italy), 2 mM L-glutamine (ECB3000D, Euroclone, Milan, Italy), and 25 mM HEPES (ECM0180D, Euroclone, Milan, Italy).
Cultures were kept in a 5% CO₂–95% air humidified incubator (Napco, model 5415, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C, fed every 2–3 days, and routinely subcultured with 0.05% trypsin–2mM EDTA solution (ECM0920D, Euroclone, Milan, Italy).

LNCaP, DU145, PC3, and 22Rv1 PCa cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in a humidified incubator in 5% CO2 at 37 °C. DU145 and PC3 cells were cultured in RPMI-1640 medium (#30-2001, ATCC) supplemented with 10% Fetal Bovine Serum (FBS), (certified, One Shot™ format, Thermo Fisher Scientific, Waltham, MA, USA); 22Rv1 cells were cultured in Dulbecco’s modified Eagle’s Medium (DMEM) with 10% FBS; LNCaP cells were cultured in RPMI 1640 with 10% FBS supplemented with 10 mM HEPES (#ECM0180D, Euroclone, Mlan, Italy) and 1 mM Sodium Pyruvate (#ECM0542D, Euroclone, Mlan, Italy). All media were supplemented with antibiotics (150 U/mL penicillin, 200 U/mL streptomycin) (#ECB3001D, Euroclone, Mlan, Italy) and 2 mM Glutamine (#ECB3000D, Euroclone, Mlan, Italy). All cell lines were routinely tested using a PCR Mycoplasma Detection Set (#6601, Takara). Cell line authentication (STR) was carried out.