Sh sy5y

The neuroblastoma cell lines CHLA-15, CHLA-20, CHLA-42, CHLA-90, CHLA-95, CHLA-171, COG-N-426 (Felix), LA-N-5, LA-N-6, NB-1643, NB-EBC1, SK-N-BE(1), SK-N-BE(2), SK-N-FI, SMS-KAN, SMS-KCR, and SMS-LHN were obtained from COG. CHP-134, IMR-32, KELLY, LA-N-1 and SH-SY5Y were obtained from the European Collection of Authenticated Cell Cultures (ECACC). GI-ME-N, NBL-S and NGP were obtained from German Collection of Microorganisms and Cell Cultures (DSMZ) and 293FT was obtained from Thermo Fisher Scientific. CHLA-15, CHLA-20, CHLA-42, CHLA-90, CHLA-95, CHLA-171, COG-N-426 (Felix), NB-1643, NB-EBC1 and NBL-S cells were cultured in IMDM (Gibco, Cat#21980032) supplemented with 20% FBS, 1% insulin-transferrin-selenium (ITS; Gibco, Cat#41400045) and 1% penicillin/streptomycin (PS). CHP-134, GI-ME-N, IMR-32, KELLY, LA-N-1, LA-N-5, LA-N-6, NGP, SK-N-BE(1), SK-N-BE(2), SK-N-FI, SMS-KAN, SMS-KCNR, and SMS-LHN cells were cultured in RPMI 1640 medium (Gibco, Cat#21875091) supplemented with 10% FBS, 1% insulin-transferrin-selenium (ITS) and 1% penicillin/streptomycin (PS). SH-SY5Y and 293FT cells were cultured in DMEM (Gibco, Cat#41966029) supplemented with 10% FBS and 1% PS. Cells were grown at 37 °C in a humidified incubator with 5% CO₂. All cells were mycoplasma-free and subjected to quarterly in-house testing using the EZ-PCR Mycoplasma Detection Kit (Geneflow, Cat#K1-0210).

Human neuroblastoma cell line SH-SY5Y was obtained from the European Collection of Authenticated Cell Cultures (ECACC) (Salisbury, UK). The cell line was authenticated by the vendors. SH-SY5Y cells were grown in Ham’s F12/MEM medium (1:1) (Sigma-Aldrich, St. Louis, MO, USA) containing 2 mM L-glutamine (Sigma-Aldrich) and 1% non-essential amino acids (NEAA) (Sigma-Aldrich). The cells were maintained at 37 °C in a humidified atmosphere of 5% CO2 in air. Cells were split every 72 h using 0.25% trypsin/EDTA (Sigma-Aldrich). After resuscitation, the cells were used for no more than 15 passages. The cells were checked for mycoplasma by using the MycoFluor Mycoplasma Detection kit (Invitrogen-Life Technologies, Monza, Italy).

Human neuron-like cell line SH-SY5Y obtained from the European Collection of Authenticated Cell Cultures (ECACC) were maintained in 75 cm² flasks containing DMEM/F12 medium (1 : 1) supplemented with 10% fetal bovine serum (FBS) and 1× antibiotic/antimycotic solution (Sigma-Aldrich). Cells were cultured in a humidified incubator set at 37°C with 5% CO₂. When cultures reached confluence, cells were trypsinized and seeded at a density of 30 × 10³ cells/cm² in 96-well culture plates. Treatments started 24 hours after seeding. All treatments were performed using 1% FBS supplemented medium. Bordo juice and wine were freeze-dried to remove water and alcohol and then dissolved in culture medium at the desired concentration (w/v). Cells were exposed to these juice and wine solutions or vehicle (culture medium).