2.2. Neuron Culture and transfection
All animal experiments were approved by the Animal Care and Use
Committee of the Institutional Animal Committee of Suzhou Institute of
Nano-Tech and Nano-Bionics, Chinese Academy of Sciences. Primary neuron
culture was performed according to the previous
article.[17] Briefly, hippocampal neurons from
embryonic day 18 (E18) Balbc rats (Cavens Lab Animal Co., Changzhou,
China) were dissected and dissociated by trypsin, the single-cell
suspension was plated on 12 mm 0.01% laminine-coated, 0.1%
poly-D-lysine pre-coated coverslips at a density of 2,400
cells/cm2. The cells were cultured in Neurobasal media
including 1% penicillin-streptomycin and 1% L-Glutamine supplemented
with 2% B27 and 1% sodium pyruvate. Neuron transfection was operated
after 5 days of culture. Each coverslip was renewed with merely
Neurobasal media 4 hours in advance, followed by liposome transfection.
The amount of vector
was
applied empirically to allow sparse transfected cells to be visualized.