2 μg/mL heparin (Sigma-Aldrich), cells were dissociated and cultured in 24-well plates on 0.1mg/mL polyornithine and Matrigel (BD Biosciences, Bedford, MA) coated glass coverslips in neural differentiation media containing the neural growth factors 1x B27 supplement (Thermo Fisher Scientific), 1μg/mL laminin (Sigma-Aldrich), and 10ng/mL of each brain-derived neurotrophic factor (BDNF, Peprotech, Rocky Hill, NJ), glial-derived neurotrophic factor (GDNF, Peprotech), and insulin-like growth factor 1 (IGF-1, Peprotech). All cells were incubated at 37° in 5% CO2.
