In-vitro separation culture method for hippocampal neurons of adult rat
A technique for separating and culturing hippocampal neurons, which is applied in the field of isolating and culturing adult rat hippocampal neurons in vitro, which can solve the problems of loss of neurotrophic factors, failure of long-term culture, and destruction of synaptic structures, etc., and achieve the effect of maintaining neuron activity
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[0040] The technical solutions of the present invention will be further described below through specific embodiments.
[0041] The present invention aims to provide a set of relatively mature solutions for the isolation and long-term in vitro culture of adult rat hippocampal neurons. The construction of flow culture system for long-term culture in vitro. Specifically include the following steps:
[0042] 1. Coated culture plate
[0043] 1) First install the inlet tube and the outlet tube of the perfusion culture plate with sterile miniature three-way pistons, and then spread a sterile circular cover glass for cell culture in each well of the culture plate, 4 pieces / well;
[0044] 2) Dissolve 1mg poly-D-lysine in 10ml sterile water to prepare 100ug / ml poly-D-lysine solution (dissolved in sterile water), pipette 1ml / well into the culture In the plate, gently rotate the culture plate, so that the liquid evenly covers the cover glass at the bottom of the plate, and coat overnig...
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