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Cancer is a global concern and the potential targeting of cancer stem cells (CSCs) has gained major attention in anticancer drug discovery research. Various natural compounds have shown promising effects in suppressing cancer, prompting scientific interest. In this study, we investigated the anti-CSC properties of schisandracaurin C, a novel compound derived from the Vietnamese plant Schisandra cauliflora. To evaluate its anti-CSC efficacy, we employed the MTT assay, flow cytometry analysis, the caspase-3 assay and a three-dimensional cell culture method, focusing on the teratocarcinoma NTERA-2 cells. Our findings revealed that schisandracaurin C inhibited the growth of NTERA-2 cells with an IC50 value of 16.61±0.13 μM, which was significantly lower than that on HEK-293A healthy embryonic kidney cells (22.72±0.02 μM) (P<0.05). Schisandracaurin C was toxic to NTERA-2 cells within three-dimensional tumorspheres (IC50: 33.47±1.45 μM). The anti-CSC effects of schisandracaurin C on NTERA-2 cells were mediated through apoptosis, as evidenced by increased caspase-3 activity. Schisandracaurin C induced cell cycle arrest at the S phase in NTERA-2 cells. These findings highlight the potential of schisandracaurin C as a promising anti-CSC compound, which warrants further mechanistic and clinical investigations. [ABSTRACT FROM AUTHOR]