Effect of glass or polystyrene substrates and cell-seeding density on dendrites of wild-type mouse hippocampal neurons. (A) DIV 12 mouse hippocampal neurons immunostained for dendrites cytoskeleton (MAP2, red) and nuclei (Hoechst, blue) plated at different seeding cell densities on glass coverslips (top lane) or polystyrene plates (bottom lane) (Scale bar = 100 µm). (B) Number of neurons per mm2 counted at DIV 12 for the different seeding cell densities. Data are expressed as mean ± SEM, n = 4 cultures per condition on glass, n = 3 cultures per condition on polystyrene. (C) Percentage of neurons (%) normalized on the number of counted viable nuclei (total cells), (D) average Total Dendritic Length (TDL) per neuron (µm), (E) average number of dendrites endpoints per neuron and (F) average neuronal soma area (µm2). Number of neurons measured ranged from 1000 for the highest seeded cell density, to 200 neurons for the lowest seeded cell density. Unpaired t-test to compare the different substrates at same cell density, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. One-way ANOVA was used to compare TDL, endpoints and soma area at different dilutions (D,E), ns = not significant difference.