The characteristics of spinal cord-evoked potentials were investigated by using an in vitro spinal cord preparation. The spinal cord isolated from adult rats was immersed in a bathing chamber filled with oxygenated artificial cerebrospinal fluid (aCSF). The spinal cord-evoked potentials elicited by the stimulation of the spinal cord were recorded by using bipolar platinum electrodes. The potentials recorded consisted of early and late negative components (N1, N2). The inhibitory effect of lowered temperature on the N1 potential was clearly demonstrated. The oxygen deprivation of aCSF showed the higher sensitivity of N2 potential to hypoxia than that of N1. It was also possible to analyze the effects of potassium and magnesium ion concentration and pH on the evoked potentials. These results showed that the pure effects of various physiological and chemical factors on the spinal cord-evoked potentials can be analyzed by this experimental model. The characteristics of spinal cord-evoked potentials were investigated by using an in vitro spinal cord preparation. The spinal cord isolated from adult rats was immersed in a bathing chamber filled with oxygenated artificial cerebrospinal fluid (aCSF). The spinal cord-evoked potentials elicited by the stimulation of the spinal cord were recorded by using bipolar platinum electrodes. The potentials recorded consisted of early and late negative components (N1, N2). The inhibitory effect of lowered temperature on the N1 potential was clearly demonstrated. The oxygen deprivation of aCSF showed the higher sensitivity of N2 potential to hypoxia than that of N1. It was also possible to analyze the effects of potassium and magnesium ion concentration and pH on the evoked potentials. These results showed that the pure effects of various physiological and chemical factors on the spinal cord-evoked potentials can be analyzed by this experimental model.