Primary murine high-grade glioma cells derived from RCAS/tv-a diffuse glioma model reprogram naïve T cells into immunosuppressive regulatory T lymphocytes.

High-grade gliomas (HGG) and glioblastomas (GBM) are the most aggressive and lethal brain tumors. The standard of care (SOC) currently includes gross safe surgical resection followed by chemoradiotherapy. The main chemotherapeutic agents are the DNA alkylating agent Temozolomide (TMZ) and adjuvants. Due to the outdated therapeutic protocols and lack of specific treatments, there is an urgent and rising need to improve our understanding on the tumor biology and to design more effective therapeutic strategies. In vitro models are essential for investigating glioma biology and testing novel therapeutic approaches. While the use of commercially available and patient-derived glioma cell lines for in vitro studies is a common practice, they exhibit several limitations including failing to maintain genetic and phenotypic diversity of primary tumors, going through genetic drift over time, and often lacking the invasive and stem-like characteristics of patient tumors. These limitations can lead to inconsistent and non-reproducible results, hampering translational research progress. In this study, we established a novel primary murine high-grade glioma cell line, isolated from an immunocompetent HGG-bearing RCAS/t-va mouse. We characterized the transcriptome and phenotype to ensure this cell line resembles the nature of high-grade gliomas and retains the ability to reprogram primary murine T lymphocytes. RNA was extracted from #4 CT2A and #4 BRCAS1 cell lines after 4 days of cell culturing, and wih less than 8 passages. Samples were processed for total RNAseq