Background: Plants rich in essential oils belong to more than 50 families, including Asteraceae, Rutaceae, and Lamiaceae. These plants are widely used in food, traditional and modern medicine, perfumery, cosmetics, and pharmaceutical industries. Approximately 3,000 types of essential oils have been identified, of which more than 300 are utilized for industrial and commercial purposes. Essential oils exhibit antimicrobial, anti-inflammatory, anticancer, cytotoxic, and antioxidant activities and are therefore widely applied in medical research. In Mongolia, the extreme climate, distinct ecological regions, and the prevalence of pastoral livestock husbandry provide a scientific basis for investigating essential oil–containing plants, their chemical composition, and biological activities. Among them, Juniperus Sabina L. is of particular theoretical and practical interest. Aim: To determine the non-toxic dose of essential oil extracted from Juniper Sabina L. on neural cells. Materials and Methods: This study was conducted at the Institute of Bio-Medicine, MNUMS. Ethical approval was obtained from the Research Ethics Committee of MNUMS (Meeting date: November 21, 2025; Protocol No. 2025-26/0301). BV2 microglial and N2a neuronal cell lines were cultured under standard conditions at 37°C in a 5% CO₂ incubator. Cells were maintained in appropriate culture media, and their growth and morphology were monitored microscopically before experimentation. Cell viability was assessed using the MTT assay. BV2 microglial cells were seeded into 96-well plates at a density of 2×10⁴ cells/well and incubated for 24 hours. Essential oil was added at concentrations ranging from 50–500 µg/mL and incubated for an additional 24 hours. After treatment, MTT reagent was added and incubated for 3 hours. The resulting formazan crystals were dissolved in dimethyl sulfoxide (DMSO), and absorbance was measured at 630 nm. Cell viability was calculated based on absorbance values to determine the non-toxic concentration of the essential oil. Result: The cytotoxic effects of the essential oil were evaluated in BV2 microglial and N2a neuronal cells using the MTT assay. In N2a neuronal cells, concentrations of 50, 100, 200, and 300 µg/mL did not significantly affect cell viability, whereas 400 and 500 µg/mL reduced cell viability, indicating cytotoxic effects. Conclusion According to MTT assay results, the essential oil of Juniperus sabina showed no cytotoxic effect on N2a neuronal cells at concentrations of 50, 100, 200, and 300 µg/mL, while cytotoxic effects were observed at 400 and 500 µg/mL. In BV2 microglial cells, concentrations of 50, 100, and 200 µg/mL were non-toxic, whereas concentrations above 300 µg/mL exhibited cytotoxic effects.