In this article,the dissociation and culture of neurons isolated from the subesophageal ganglion(SUB) of the Ornithoctonus huwena are described.The basic electrophysiological properties of voltage-gated Na⁺,K⁺ and Ca²⁺ channels on the cultured neurons were studied by means of whole-cell patch-clamp technique.The culture medium used for the nerve cells contained(g/L): glucose 0.7,fructose 0.4,succinic acid 0.06,imidazole 0.06,L-15 13.7,Hepes 2.38,yeast extract 2.8,lactalbumin 2.5,penicillin 200 IU/ml,streptomycin 200 mg/ml,bovin calf serum 15%;pH 6.8.The suitable culture was 27±2℃ for 2-4 h.Most cells were in good condition and above 90% cells survived in the cell culture dishes.The shape of the soma of the nerve cell was in an ellipse and that of neural cell appreared like a spoon,with a single axon.The size of these cells varied from 10 to 30 μm.Whole-cell patch-clamp showed high-voltage-activated(HVA) calcium currents and two types of outward potassium currents including delayed rectifier potassium currents and rapid outward potassium currents on spider neurons.The potassium currents could be inhibited by TEA-Cl and 4-AP.Sometimes,small voltage-gated sodium currents were also recorded in the experiment.