Abstract TTFields are alternating electric fields of low intensity (1-3 V/cm) and intermediate frequency (100-500 kHz), which are effective and approved for the treatment of glioblastoma (GBM) using 200 kHz frequency. However, there is a lack of ex vivo models to evaluate effects on patients’ tumor biology or to screen patients for treatment efficacy. Therefore, we adapted patient-derived three-dimensional GBM tissue culture models to be compatible with TTFields application and recently published the feasibility of such an approach (Nickl, et al., 2022, doi: 10.3390/cancers14215177). Here, we applied one of those models, i.e. tumor-organoids cultured as microtumors on murine organotypic hippocampal slice cultures (OHSCs), to additional brain tumor entities, namely a sample of an anaplastic ependymoma (AE) patient and an oligodendroglioma patient. Organoids were generated from fresh intra-operatively obtained tumor tissue and cultured for 2 weeks. OHSCs were prepared by slicing the brains of mice 5-8 days postpartum to sections with a thickness of 350 µm using a vibratome, and culturing them for 2 weeks as well. Subsequently, organoids were placed onto the OHSCs. The inovitro™ laboratory research system was used for TTFields administration at 200 kHz and 1.5 V/cm for 72 h. Microtumor growth was evaluated on fluorescence images. Viable organoids formed from the GBM, AE and oligodendroglioma sample and grew to microtumors when placed onto OHSCs. Application of TTFields at 200 kHz led to a significant decrease of microtumor size of the GBM and AE (both p<0.0001), but not the oligodendroglioma sample. This proof-of-principle investigation proved that the application of patient-derived organoids cultured on OHSCs is feasible to investigate the effects of TTFields on different kinds of brain tumors. To our knowledge, this is the first evaluation of TTFields efficacy on patient derived AE and oligodendroglioma tissue cultures. While TTFields at 200 kHz led to a decrease in the microtumor size of the AE sample, the non-responsiveness of the oligodendroglioma sample may be due to different inter-patient sensitivity to TTFields or a suboptimal TTFields frequency. Citation Format: Vera Nickl, Ellina Schulz, Ellaine Salvador, Laureen Trautmann, Leopold Diener, Almuth F. Kessler, Camelia M. Monoranu, Ralf-Ingo Ernestus, Mario Löhr, Carsten Hagemann. Evaluation of tumor treating fields (TTFields) effects at 200 kHz on a glioblastoma, an anaplastic ependymoma and an oligodendroglioma sample in a patient-derived ex vivo organoid model. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4573.