The purpose of this study was to develop, assess, and validate a custom 32-channel system to analyze the electrical properties of 3-D artificial heart muscle (3D-AHM). In this study, neonatal rat cardiac cells were cultured in a fibrin gel to drive the formation of 3D-AHM. Once the tissues were fully formed, the customized electrocardiogram (EKG) sensing system was used to obtain the different electrophysiological characteristics of the muscle constructs. Additionally, this system was used to evaluate the electrical properties of native rat hearts, for comparison to the fabricated tissues and native values found in the literature. Histological evaluation showed extensive cellularization and cardiac tissue formation. EKG data analysis yielded time delays between the signals ranging from 0 to 7 ms. Optical maps exhibited slight trends in impulse propagation throughout the fabricated tissue. Conduction velocities were calculated longitudinally at 277.81 cm/s, transversely at 300.79 cm/s, and diagonally at 285.68 cm/s for 3D-AHM. The QRS complex exhibited an R-wave amplitude of 438.42 ± 36.96 μV and an average duration of 317.5 ± 16.5 ms for the tissue constructs. The data collected in this study provide a clearer picture about the intrinsic properties of the 3D-AHM while proving our system's efficacy for EKG data procurement. To achieve a viable and permanent solution, the bioengineered heart muscle must physiologically resemble native heart tissue as well as mimic its electrical properties for proper contractile function. This study allows us to monitor such properties and assess the necessary changes that will improve construct development and function.