Parylene is used as encapsulating material for medical devices due to its excellent biocompatibility and insulativity. Its performance as the insulating polymer of implantable neural interfaces has been studied in electrolyte solutions and in vivo. Biological tissue in vitro, as a potential environment for characterization and application, is convenient to access in the fabrication lab of polymer and neural electrodes, but there has been little study investigating the behaviors of Parylene in the tissue in vitro. Here, we investigated the electrochemical impedance behaviors of Parylene C polymer coating both in normal saline and in a chilled pig brain in vitro by performing electrochemical impedance spectroscopy (EIS) measurements of platinum (Pt) wire neural electrodes. The electrochemical impedance at the representative frequencies is discussed, which helps to construct the equivalent circuit model. Statistical analysis of fitted parameters of the equivalent circuit model showed good reliability of Parylene C as an insulating polymer in both electrolyte models. The electrochemical impedance measured in pig brain in vitro shows marked differences from that of saline.