Battery impedance is essential to the management of lithium-ion batteries for electric vehicles (EVs), and impedance characterization can help to monitor and predict the battery states. Many studies have been undertaken to investigate impedance characterization and the factors that influence impedance. However, few studies regarding the influence of the internal temperature gradient, which is caused by heat generation during operation, have been presented. We have comprehensively studied the influence of the internal temperature gradient on impedance characterization and the modeling of battery impedance, and have proposed a discretization model to capture battery impedance characterization considering the temperature gradient. Several experiments, including experiments with artificial temperature gradients, are designed and implemented to study the influence of the internal temperature gradient on battery impedance. Based on the experimental results, the parameters of the non-linear impedance model are obtained, and the relationship between the parameters and temperature is further established. The experimental results show that the temperature gradient will influence battery impedance and the temperature distribution can be considered to be approximately linear. The verification results indicate that the proposed discretization model has a good performance and can be used to describe the actual characterization of the battery with an internal temperature gradient.