Sh sy5y human neuroblastoma cell line

Experiments were carried out using the SH-SY5Y human neuroblastoma cell line originally from ATCC (Rockville, MD, USA). Their differentiation was performed with 10 µM retinoic acid (RA; Sigma–Aldrich, Milan, Italy) in MEM/Ham’s F12 medium supplemented for 5 days [17 (link)]. All reagents were from Gibco (Life Technologies, Monza, Italy). The stock solution of MJe 400 mg/mL was prepared in DMSO that was employed, upon further dilution in culture medium, to obtain the working concentrations. The same percentages of DMSO present in these dilutions served as vehicle controls that were tested in each of the following experiments to confirm that no effect was induced by the solvent (data not shown).

HT22 mouse hippocampal neuronal cell line was purchased from Sigma (SCC129, St. Louis, MO, USA). SH-SY5Y human neuroblastoma cell line was purchased from ATCC (CRL-2266, Manassas, VA, USA). Cells were cultured in DMEM medium (11965092, Life Technology, Rockford, IL, USA) supplemented with 10% fetal calf serum (30-2030, ATCC), 100 U/mL penicillin G sodium (P3032, Sigma), and 100 μg/mL streptomycin sulfate (11860038, Thermo Fisher). The cells were maintained at 37 °C in a humidified atmosphere with 5% CO₂. The medium was changed every two days. HT22 and SH-SY5Y cells were used within 8 generations.

The SH-SY5Y human neuroblastoma cell line was obtained from ATCC (Manassas, VA) and was cultured and maintained as described earlier [26 (link)]. For passaging and for all experiments, the cells were detached using 0.25 % trypsin /1 % EDTA, resuspended in complete medium, counted (Countess, Invitrogen), and incubated in a 95 % air/5 % CO₂ humidified incubator.

Experiments were carried out using the SH-SY5Y human neuroblastoma cell line (originally from ATCC, Rockville, MD, USA). Cells were differentiated in MEM/Ham’s F12 medium supplemented with 10-µM retinoic acid (RA; Sigma–Aldrich, Milan, Italy) for 5 days as reported by Condello et al. [28 (link)]. All reagents were from Gibco (Life Technologies, Monza, Italy).

The SH-SY5Y human neuroblastoma cell line was obtained from ATCC (Manassas, VA) and was cultured and maintained as described earlier [54 (link)]. For passaging and for all experiments, the cells were detached using 0.25 % trypsin /1 % EDTA, re-suspended in complete medium, counted (Countess, Invitrogen), and incubated in a 95 % air/5 % CO₂ humidified incubator.

SH-SY5Y human neuroblastoma cell line was purchased (ATCC, USA) and they were cultured with Dulbecco’s Modified Eagles Medium (DMEM) containing 10% (v/v) heat-inactivated fetal bovine serum (Gibco, Gaithersburg, MD, USA) and 100U penicillin–streptomycin (Gibco).
Cells were seeded in a 96-well plate at a density of 10,000cells/well; after 24 h, 4 different Aβ1-42 (Abcam, Cat No: 120301; Cambridge, UK) concentrations (1.25µM, 2.5 µM, 5 µM, and10 µM) were applied in order to find the effective toxic dose. Aβ1–42 was dissolved in 1% (v/v) NH4OH (Larson, 2016). Lactate dehydrogenase (LDH) assay (Roche, Cat No: 11644793001; Mannheim, Germany) was performed for cell viability measurements. Briefly, the working solution of the LDH assay was prepared according to the manufacturer’s instructions and was incubated in complete darkness for 15 min at room temperature. The medium in which our cells were cultured was put into a new 96-well plate in exactly the same order as the original plate. Next, 100 µL of LDH working solution was added to each well containing the cell culture medium to produce a final volume of 200 μL. At 492 nm, absorbance values in each well were measured in a microplate reader (Chromate Manager 4300, Awareness Technology, Palm City, FL, USA).