Research Paper

Patient glioma-derived stem cells formed neurosphere-like colonies and demonstrated characteristics of NSCs. (A) Morphology of typical primary neuorspheres derived from GBM tissues. The scale bar in high-magnification images (cyan arrow pointed region in low-magnification image) represent 50 μm. (B) The neurosphere formation time on different GSCs. Five primary and paired recurrent GBM tissues were obtained and cultured for at least 7 d. Neurospheres from 4 recurrent and 3 primary GBMs were formed. The recurrent GSCs showed higher formation ability compared with the primary GSCs (*: P < 0.05 and **: P < 0.01). (C) Immunofluorescent staining of primary tumor spheres with stem cell markers (CD133 [green]) and Nestin [red]) confirmed stemness of neurospheres. (D) The neurospheres were cultured in differentiation medium for 7 d, and differentiation markers (GFAP [green] and beta-tubulin III [red]) were immunofluorescence-stained. The obtained neurospheres maintained the typical multipotency ability. The scale bar in high-magnification images (white arrow in low-magnification images) represents 20 μm.