目的：分离培养人脑胶质瘤干细胞样细胞（glioma stem-like cell，GSLC），研究其体外侵袭力。方法：选取中国医科大学第一医院神经外科2008年10月至2009年1月间住院患者手术切除的脑胶质瘤组织8例，以无血清成球培养法培养胶质瘤细胞球；免疫细胞化学实验检测其CD133的表达；荧光免疫显微镜观察其分化后胶质细胞标志物GFAP和神经元标志物TU-20的表达；matrigel侵袭实验检测其侵袭力，并与原代脑胶质瘤细胞进行比较。结果：成功分离培养出人脑胶质瘤细胞球细胞，该细胞表达干细胞标志物CD133；能自我更新与增殖；诱导分化后，GFAP和TU-20均为阳性表达，提示其为GSLC。胶质瘤细胞球细胞侵袭细胞数显著多于原代胶质瘤细胞\[（261.23±87.20）vs（116.08±63.88）个，P<0.01\]；此外，胶质瘤细胞球细胞穿过matrigel胶后可再次聚集成球状生长。结论：成功分离培养人脑胶质瘤组织中的GSLC，其体外具有较高的侵袭力，可能参与脑胶质瘤的侵袭和转移。

Objective:To isolate and culture human brain glioma stem-like cells (GSLCs), and to determine the invasiveness of GSLC in vitro. Methods: Glioma cell spheres were serum free cultured from 8 surgical specimens of glioma inpatients from October 2008 to January 2009 in the Department of Neurosurgery, First Hospital of China Medical University. CD133 expression was identified by immunocytochemistry assay, and the expressions of GFAP and TU-20 in differentiated glioma cell spheres were detected under an immunofluorescence microscope. With matrigel invasion assay, invasiveness of glioma sphere cells was determined and compared with that of primary glioma cells. Results: Glioma cell spheres were cultured successfully. These cells were proved to express stem cell marker CD133, capable of renewal and proliferation, and can differentiate to glia and neurons with positive expressions of GFAP and TU-20, indicating the characteristics of GSLC. The invasiveness of glioma spheres cells in vitro was higher than that of primary glioma cells (261.23±87.20 vs 116.08±63.88, P<0.01). Moreover, glioma spheres cells preferred to aggregate and reform new spheres after travelling through the matrigel. Conclusion:〗GSLCs are successfully cultured from human glioma tissues, which show higher invasiveness in vitro. GSLC may participate in glioma invasion and metastasis.

国家自然科学基金（No. 81000565）