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Objective: The new glioma cell line SHG-139 was established and its phenotype, tumorigenicity, pathological characteristics, derived stem cells SHG139S were studied., Methods: Immunohistochemistry was used to assess expressions in the patient and mouse tumor tissues, SHG-139 and SHG-139S. Primary SHG-139 culture was performed, cell proliferation, cell cycle and genetic characteristics were assessed. MiRNA (Micro RNA) and LncRNA (Long non-coding RNA) microarray was performed., Results: We found that the glioma tissue was positive for A2B5 (Glial precursors ganglioside), GFAP (Glial fibrillary acidic protein), S-100 (Acid calcium bingding protein), VEGF (Vascular endothelial growth factor), VEGFR (Vascular endothelial growth factor receptor) and negative for Ki-67 (Nuclcar- associated antigen). SHG-139 proliferated significantly within 24h; its total number of chromosomes was 68; ratios of SHG-139 and SHG-139S cells in G1 phase were highest. SHG-139 cells were positive for A2B5, GalC (Galactocerebrosides), GFAP, S-100 and Vimentin, while SHG-139S cells were positive for A2B5, Nestin, and NG2 (Neuron-glia antigen2), and negative for Vimentin and IDHR132H (Isocitrate dehydrogenase); cells rarely stained for CD133 (Cluster of differentiation133). SHG-139 intracranial xenografts expressed GFAP, but no overt oligodendroglioma was observed. In SHG-139S xenografts, GFAP and S-100 were expressed, while CD133 was not detected; a few A2B5(+) cells were found at tumor edges, and typical oligodendroglioma were obtained. In addition, SHG-139S xenograft tumors were more aggressive than those of SHG-139. Anti-mouse CD31 (Cluster of differentiation31) staining revealed murine vessels at the border between xenograft tumor and normal brain tissue; Anti-human CD34 (Cluster of differentiation34) staining was negative. Biochip technology of SHG139S showed several miRNA and lncRNA were differently expressed in SHG139 and SHG139S., Conclusions: SHG-139 was an astroglioma cell line which yielded stem cells SHG-139S. SHG-139S cells constituted an A2B5(+)/CD133(-) GSC subgroup.