We expanded GSCs from glioma LN18 cells in serum-free medium with growth factors and confirmed the higher expression of pluripotency markers (NANOG, SOX2, CD133) in GSCs-enriched spheres, as previously shown (Ciechomska et al., 2016; Kijewska et al., 2017). The gene discussed is PROM1; the disease is central nervous system cancer.