Objective　　　To explore the role of Hedgehog pathway in endothelial cells promoting self-renewal of glioma stem cell (GSC).　　　 Methods　　　GSC derived from glioblastoma cell line GL261 and brain microvessel endothelial cell line b.END3 were used. Transwell co-culture system

limit dilution assay

real-time PCR

Western blotting

xenograft experiment and gene knock-down assay were applied to determine the self-renewal

tumorigenic ability and gene expression of Hedgehog pathway in GSC spheres.　　　 Results　　　(1) More and larger tumor spheres were formed by GSC after co-culture with endothelial cells (P0.05)

especially under a low cell concentration of 5 cells per well (24.3% vs 11.3%) and 10 cells per well (39.4% vs 25.8%). (2) Hedgehog pathway related genes including Gli1

Oligo2 and Bmi1 were up-regulated in the co-cultured GSC spheres (P0.05). (3) Larger xenografts were generated by GSC spheres mixed with endothelial cells

which also resulted in tumor rupture and model death (P0.05). (4) The phenomenon above nearly disappeared when Hedgehog pathway had been inhibited by Smo gene knockdown.　　　 Conclusion　　　Endothelial cells promote self-renewal of GSC through activating Hedgehog pathway.