2.8. Cell Culture

Monolayer-adherent HEK-293T cells (transformed human embryonic kidney cells) and SY5Y-SH cells (bone-marrow-derived human neuroblastoma cells) were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco by Thermo Fisher Scientific, USA) supplemented with 10% (vol/vol) fetal bovine serum (FBS) (Thermo Fisher Scientific, USA) and 0.01% (vol/vol) pen–strep solution ×10 (Biological Industries, Israel).

Cells were incubated at 37 °C in a 5% CO₂ atmosphere. Before use, each cell line was confirmed to have no mycoplasma contamination using the EZ-PCR Mycoplasma Detection Kit (Biological Industries, Israel).

Prior to each experiment, the cells were dissociated using trypsin 0.25% EDTA (Thermo Fisher Scientific, Israel), stained with trypan blue (Biological Industries, Israel), and counted using the Countess^TM II Automated Cell Counter (Invitrogen at Thermo Fisher Scientific, USA).