Expression of neural stem cell and glial progenitor surface markers in low- and high-grade glioma specimens analyzed.

Freshly prepared or thawed glioma cells were first incubated with non-specific mouse IgG1 mAb, cells were subsequently incubated with APC conjugated anti-CD45 mAb in combination with FITC conjugated anti-CD44, or anti-CD24 mAbs or PE conjugated anti-PDGFRα, anti-EGFR, anti-CD133 mAbs. For A2B5 and O4 staining, cells were incubated with unconjugated A2B5 or O4 mAb respectively and subsequently stained with PE conjugated rat anti-mouse IgM. Cells negatively stained with 7-AAD were analyzed for cell surface marker expression. Data shown are the percentages of CD45 negative cells positively stained for indicated cell surface markers.
*: secondary GBM. UD: undetectable; ND: not done. PNET: primitive neuroectodermal tumor.

将新鲜制备或复苏解冻的胶质瘤细胞，首先以非特异性小鼠IgG1单克隆抗体（mAb）进行孵育；随后将细胞与别藻蓝蛋白（APC）标记的抗CD45单克隆抗体，分别联合异硫氰酸荧光素（FITC）标记的抗CD44或抗CD24单克隆抗体，或联合藻红蛋白（PE）标记的抗PDGFRα、抗EGFR、抗CD133单克隆抗体共同孵育。针对A2B5与O4染色实验，分别将细胞与未标记的A2B5单克隆抗体、O4单克隆抗体孵育，之后使用藻红蛋白（PE）标记的大鼠抗小鼠IgM进行二抗染色。对经7-氨基放线菌素D（7-AAD）阴性染色的细胞开展细胞表面标志物表达分析。本次展示的数据为CD45阴性细胞中，各指定细胞表面标志物阳性染色的细胞占比。
*：继发性胶质母细胞瘤（secondary GBM）；UD：未检出（undetectable）；ND：未完成（not done）；PNET：原始神经外胚层肿瘤（primitive neuroectodermal tumor）。