Expression profiles of mouse glioma-initiating cells.

To identify factors involved in glioma-initiating cells (GICs), we compared gene expressions between GIC-like cells and non-GICs. p53-deficient Neural stem cells (NSCs), oligodendrocyte precursor cells (OPCs), and astrocytes (ASTs) were transfected with pCMS-EGFP-HRasL61 and pBabe-neo by electroporation and cultured in 0.5 mg/ml neomycin. The GFP-positive stable NSCs (NSCL61s), OPCs (OPCL61s) and ASTs (ASTL61s) were purified by flow cytometry. Total RNA was prepared using a RNeasy Mini Kit (QIAGEN) and used for the expression analysis.为鉴定参与胶质瘤起始细胞（glioma-initiating cells, GICs）生物学过程的调控因子，本研究对比了类GIC细胞与非GIC细胞的基因表达谱。我们采用电转染法将pCMS-EGFP-HRasL61与pBabe-neo载体导入p53缺陷型神经干细胞（Neural stem cells, NSCs）、少突胶质前体细胞（oligodendrocyte precursor cells, OPCs）及星形胶质细胞（astrocytes, ASTs），随后将细胞置于含0.5 mg/ml新霉素的培养基中培养。通过流式细胞术纯化获得GFP阳性的稳定转染神经干细胞（NSCL61s）、少突胶质前体细胞（OPCL61s）与星形胶质细胞（ASTL61s）。采用RNeasy迷你试剂盒（QIAGEN）提取总RNA，用于后续基因表达分析。