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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 glioma stem-like cells (GSLC) and investigate their in vitro invasiveness. Methods: Eight cases of glioma were removed from the hospitalized patients in Department of Neurosurgery, First Hospital of China Medical University from October 2008 to January 2009, and glioma cells were cultured in serum-free culture. Immune cells The expression of CD133 was detected by chemical assay, the glial cell marker GFAP and the neuronal marker TU-20 were observed by fluorescence immunohistochemistry. The invasiveness of matrigel was tested by invasive immunocytochemistry. Compare Results: The human glioma cells were successfully isolated and cultured. The cells expressed stem cell marker CD133, self-renewing and proliferating. GFAP and TU-20 were positive after induced differentiation, suggesting that they were GSLC. The number of glioma cells invaded by glioma cells was significantly more than that in primary glioma cells [(261.23 ± 87.20) vs (116.08 ± 63.88), P <0.01]. In addition, glioma cells passed through matrigel Can be gathered again into a spherical growth. CONCLUSION: The successful isolation and culture of GSLC in human glioma tissue has high invasiveness in vitro and may be involved in the invasion and metastasis of glioma.