Primary Mouse Cortical Neuron Culture

Wildtype mice were mated and at e15 the females were euthanized and the embryos were removed. The females were not further used. The embryos were decapitated and cortex tissue was dissected and used for primary cultures. Culture set-up is previously described in Perland et al. (2016) (link).Briefly, the cortex samples were pooled, washes in PBS-Glucose and dissociated using papain (Thermo Fisher Scientific) and DNAse (Thermo Fisher Scientific) for 30 min. Thereafter, mechanical dissociation was performed by pipetting before filtering the cell solution through a cell strainer (Thermo Fisher Scientific). The cells were diluted in plating media consisting of DMEM:F12 (Gibco, Invitrogen) supplemented with 2 mM GlutaMax, 1 mM Na-Pyruvate, 10% FBS and 1% Pen strep, all supplied from Invitrogen, and plated on Poly-L-Lysine (Sigma) coated coverslips (12 mm, #1.5) (Menzel-Gläser, Thermo Fischer Scientific) in 24-well plates (Nonclone delta, Thermo Fischer Scientific) or on Poly-L-Lysine coated 6-well plates (Nunclone delta, Thermo Fischer Scientific). The cell was incubated at 37°C, 5% CO₂ for 3 h before media change to NeurobasalA media (Gibco, Invitrogen) supplemented with 2 mM Glutamax, 1 mM Na-Pyruvate, 1 % Pen-strep and 1x B27 Supplement. 75% of the media was changed every third day and the cells allowed to grow for 9 days.