Culturing Neural Stem Cells with Amyloid-beta Treatment

A neural stem cell (NSC) line (NMW7) was generated from a mouse fetal brain, as described previously [57 (link)]. The cells were cultured with medium containing high glucose DMEM (Wako Pure Chemicals, Richmond, VA, USA): F12 ham (Wako) 1:1, bFGF (PeproTech, Rocky Hill, NJ, USA), 20 ng/mL, EGF (peproTech), 20 ng/mL, N2 supplement (ThermoFisher, Waltham, MA, USA), and 2% FBS (Gibco, Invitrogen) in an attached culture condition. The NSC was sub-cultured every 48 h. During stimulation, high glucose DMEM medium containing 0.2% FBS, with or without indicated concentrations of Aβ_1–42 (Peptide Institute, Osaka, Japan), was used. Aβ_1–42 was added to the culture as a monomer, and the stimulations were continued for the indicated times.