Abstract Background: Tumor Treating Fields (TTFields) are an approved anti-neoplastic treatment modality delivered via application of low intensity, intermediate frequency, alternating electric fields. The electrical properties of cells (such as permittivity and conductivity) determine the optimal frequency of TTFields that incurs the highest reduction in cell counts. Currently, there are no predictive markers for determining TTFields response or the optimal frequency to be applied for individual patient. The goal of this study is to determine the correlation between electrical properties of cells and TTFields’s optimal frequency and sensitivity. The 3DEP reader (LabTech) determines the electrical properties of cells, including permittivity and conductivity, by using Dielectrophoresis (DEP) force. DEP is a physical effect that generates a force on polarizable particles experiencing a non-homogeneous electric field and can therefore be used as a technique to analyse the way cells move within electric fields at different frequencies. Methods: The baseline electrical properties (permittivity and conductivity) of 17 cell lines from different tumor types were determined using the 3DEP reader (LabTech). The curves were analyzed using 2-way ANOVA. The optimal TTFields frequency for each cell line was determined by testing the cytotoxic effect of TTFields at various frequencies using the inovitro system. The electrical properties of cells were compared with the optimal TTFields frequency and sensitivity of each cell line. Results: The results demonstrate significant differences (p<0.001) between the lower frequency range of the 3DEP curves that corresponds to membrane capacitance of cells with TTFields optimal frequency of 150kHz (9 cell lines) and cells with TTFields optimal frequency of 200kHz (8 cell lines). Membrane capacitance was also a good predictor for TTFields sensitivity based on the differences in the curves in the low frequency range. Conclusions: The results presented in this study demonstrate that cell membrane capacitance correlates with TTFields optimal frequency and sensitivity. Based on the above, there is a strong rational to further explore the potential of measuring the electrical properties of cells as a predictive marker to help determine which patient will respond better to TTFields and the optimal TTFields frequency to be applied for each patient. Citation Format: Moshe Giladi, Einav Zeevi, Karnit Gotlib, Cornelia Wenger, Ariel Naveh, Zeev Bomzon, Eilon D. Kirson, Uri Weinberg, Adrian Kinzel, Yoram Palti. Testing the electrical properties of different cell lines using 3DEP reader and compare to TTFields response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2168.