Cancer stem cells are a subpopulation of tumor cells with the ability of self-renewal and are thought to drive tumor initiation and growth. Cancer stem cells are resistant to conventional therapy which poses major problem in treatment. Vitamin C is thought to have different antitumor mechanisms that differ depending on the type of tumor. Apart from disrupting the redox state in cells and causing oxidative stress, vitamin C also changes the metabolic profile of the cell. In this work, the effect of the oxidized form of vitamin C, dehydroascorbic acid (DHA), on three sarcoma stem cell types was investigated. Millimolar concentrations of DHA in medium with and without basic FGF decreased the viability of all treated cell lines. This work also demonstrated that treatments with DHA alter the expression of genes important for glycolysis and oxidative phosphorylation. Effects of DHA treatment on change of glyceraldehyde-3-phosphate dehydrogenase protein expression were also shown. However, sarcomas are a very heterogeneous group of tumors, so three types of stem cells showed different responses to DHA treatment. General response to treatment was more positive in chondrosarcoma and osteosarcoma stem cells than in rhabdomyosarcoma stem cells. DHA has shown potential as a modulator of sarcoma stem cell metabolism, but for further development as an anti-tumor therapy, the specific background of the tumor must be considered.