Primary microglial cell/injured neuron co-culture system and construction method and application thereof

[0045] Primary microglia / injured neuron co-culture system, the system includes primary microglial cells and injured neurons, wherein the injured neurons are phagocytized by primary microglial cells; the system is In vitro co-culture system.

[0046] The construction method of the primary microglia / injured neuron co-culture system comprises the following steps:

[0047] Step 1. Isolation and culture of primary microglia

[0048] Take newborn SD rats, disinfect them with 75% alcohol one by one for 30 seconds, decapitate and take out the brain and brainstem completely; under a stereomicroscope, peel off the vascular membrane, remove the brainstem and hippocampus, cut up the cerebral cortex, and temporarily store it in 0°C precooling DMEM high-sugar+10% FBS medium; cut the brain cortex into pieces, centrifuge to remove the supernatant, add 0.125% trypsin 5mL to resuspend, transfer to a 10cm culture dish, and store at 37°C, 5% CO2 Digest in an incubator for 15 minutes; add 1 mL o

[0060] The primary microglia / injured neuron co-culture system described in Example 1 is applied to the evaluation of drug efficacy for promoting nerve regeneration, including the following steps:

[0061] Step 1. Experimental Group Intervention

[0062] The traditional Chinese medicine compound "Ji Suikang" (crude drug amount 20g / kg) that promotes nerve regeneration was given to healthy SD male rats orally to prepare drug-containing serum, and rats were given the same amount of normal saline to prepare blank serum as a control.

[0063] The primary microglia / damaged neuron co-culture system was divided into three drug-containing serum groups: 2.5% (low), 5% (medium), and 10% (high), and a 10% blank serum group (negative control group) was set at the same time. ), 2ug / mL LPS+10% blank serum group (positive control group), recovered primary cultured microglial cells, added drug-containing serum or blank serum according to the grouping conditions to prepare DMEM / F12+B27 co