Experimental Materials:
1. The sciatic nerve of the 2d rat after birth;
2. 1×PBS containing no Ca 2+ and Mg 2+ , adding 200,000 IU/L penicillin, 200 mg/L streptomycin, pH 7.2;
3. Digestive juice: a mixed digest of 0.25% trypsin and 0.03% collagenase;
4. Culture medium a: DMEM culture solution, adding 0.02-0.025 mol / L HEPES, pH 7.2;
5. Culture medium b: DMEM, adding 10% FBS;
6. Culture equipment: ophthalmic scissors, ophthalmic sputum, nylon mesh with a mesh aperture of 60 μm;
experimental method:
1. Sterilize the animal with 70% ethanol, decapitate, take the sciatic nerve, and put it into the pre-cooled culture solution a;
2. Move the sciatic nerve into the mixed digestive juice and shake it for 15 min in a 37 ° C water bath;
3. Change the fresh digestive juice and digest again for 15 min;
4. Aspirate the digestive juice, wash twice with the culture solution b, then add the fresh culture solution b, and then repeatedly blow with a pipette. Using a nylon mesh having a pore size of 60 μm, the filtrate was collected and centrifuged at 2000 r/min for 10 min;
5. Aspirate the supernatant, suspend the pellet with the culture solution b, and plant the cells in a culture flask or a petri dish, and let stand for 24 hours;
6. Add 10-5mol/L cytarabine and continue to culture for 48-72h. Passaging when the cells grow to form a monolayer;
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