中文摘要：

目的：从人胶质母细胞瘤SC 326和SC 189细胞株分离培养干细胞,研究其自我更新、增殖和分化等生物学特性。方法：用无血清培养基培养人胶质母细胞瘤SC 326和SC 189细胞,检测细胞形成神经球的能力。对培养3代以上的细胞（GSC）进行神经球细胞单细胞克隆形成率实验;用第3代和第7代的细胞评价神经球单细胞克隆形成率;免疫荧光染色法检测神经球细胞的干细胞标记物表达及多向分化能力。结果：无血清培养基培养24h时SC 326、SC 189形成神经球样细胞团,1周时神经球内细胞可达数百个,3代内神经球细胞SC 326的增殖率为（2.10±4.33）%、（4.03±4.14）%和（6.91±5.12）%,SC 189为（12.79±2.32）%、（15.78±3.25）%和（37.91±4.58）%;单细胞克隆形成率GSC 326和GSC 189分别为5.56%和8.33%,次级单细胞克隆形成率分别为9.89%和14.58%;GSC 326P3和P7神经球克隆形成率最大值为（12.67±2.86）%和（20.44±1.73）%,GSC 189P3和P7神经球克隆形成率最大值为（32.00±1.00）%和（42.67±5.03）%。免疫荧光染色,干细胞表面标记物CD133和神经巢蛋白呈强阳性表达,分化标记物胶质纤维酸性蛋白（GFAP）和神经元特异性类β-微管蛋白Ⅲ（β-TubulinⅢ）表达极弱。分化诱导后分化标记物GFAP和β-TubulinⅢ的表达呈阳性。结论：早期原代胶质母细胞瘤细胞株中存在少量具有自我更新、增殖和多向分化能力的细胞,这些细胞具有明显的干细胞特性。

英文摘要：

Objective To isolate and culture the stem cells from human glioblastoma SC 326 and SC 189 cells and to investigate the biological characteristics such as self-renewal,proliferation,and differentiation.Methods The human glioblastoma SC 326 and SC 189 cells were cultured in serum-free media,the capability of forming neurospheres was detected.Clone forming rate assay was performed to assess the capacity of self-renewal and clonogenic potential of the cells.Immunofluorescence staining method was used to study the expressions of surface markers of stem cells and the multi-differentiation function of neurospheres.Results The cells isolated from SC 326 and SC 189 cultured in serum-free media for 24 h had strong characteristics of self-renewal,proliferation,and neurosphere reformation.The proliferation rates of SC 326 were（2.10±4.33）%,（4.03±4.14）%,and（6.91±5.12）%;and those of SC 189 were（12.79±2.32）%,（15.78±3.25）%,and（37.91±4.58）% in the first three generations;the rates of neurospheres formation of GSC 326 and GSC 189 single cells were 5.56% and 8.33%,and the rates of sub-spheres formation were 9.89% and 14.58%;the maximum clone formation rates of GSC 326 P3 and P7 neurospheres were（12.67±2.86）% and（20.44±1.73）%,and those of GSC 189 P3 and P7 were（32.00±1.00）% and（42.67±5.03）%.Immunofluorescence staining showed that the neuron stem cells marker CD133 and nestin positively expressed;and the glial fibrillary acidic protein（GFAP） and neuron-specific class β-tubulin Ⅲ（β-Tubulin Ⅲ） weekly expressed.After treated with differentiated media,the expressions of differentiation markers β-Tubulin Ⅲ and GFAP were positive.Conclusion A few cells existed in glioblastoma SC 326 and SC 189 cell lines in the primary and early stage have the capacities of self-renewal,proliferation and multi-differentiation and the cells present definite features of stem cells.