ReNcell VM human neural precursor (ReN) cells were purchased from EMD Millipore (Billerica, MA, USA). The cells were plated onto BD Matrigel (BD Biosciences, San Jose, CA, USA)-coated T25 cell culture flask (BD Biosciences, San Jose, CA, USA) and maintained in DMEM/F12 (Life Technologies, Grand Island, NY, USA) media supplemented with 2 μg/ml Heparin (Stemcell Technologies, Vancouver, Canada), 2% (v/v) B27 neural supplement (Life Technologies, Grand Island, NY, USA), 20 μg/ml EGF (Sigma-Aldrich, St. Louis, MO, USA), 20 μg/ml bFGF (Stemgent, Cambridge, MA, USA), and 1% (v/v) Penicillin/Streptomycin/Amphotericin b solution (Lonza, Hopkinton, MA, USA) in CO2 cell culture incubator. The cell culture media were changed every 3 days until the cells were confluent. For 2D neuronal/glial differentiation, the cells were plated onto either Matrigel-coated 24-well or 6-well plates with DMEM/F12 differentiation media supplemented with 2 μg/ml Heparin, 2% (v/v) B27 neural supplement, and 1% (v/v) Penicillin/Streptomycin/Amphotericin b solution without growth factors. A half volume of the differentiation media was changed every 3 days for 3–7 weeks. DAPT, Compound E and BACE inhibitor IV were purchased from EMD Millipore, N-Lauroylsarcosine (Sarkosyl)
