多种胶质瘤细胞系中获取胶质瘤干细胞方法的研究

Acquisition of Glioma Stem Cells from Different Glioma Cell Lines

目的:进一步证明胶质瘤干细胞是广泛存在的,并寻找一种简洁的方法从不同胶质瘤细胞系中提取肿瘤干细胞。方法:将胶质瘤细胞以合适的密度接种于96孔板中,获取胶质瘤干细胞,并通过检测其自我更新能力、多向分化能力、成瘤能力及胶质瘤干细胞标记物的表达情况对其进行鉴定。结果:多种细胞系中均成功获取了胶质瘤干细胞。并且这细胞球表达神经干细胞的标志物,不表达神经细胞分化标志物,同时又有多向分化的能力,仅5000个细胞就可以在裸鼠颅内成瘤。结论:我们的研究结果表明胶质瘤干细胞是广泛存在的,并为以后进一步研究胶质瘤干细胞的特性及靶向胶质瘤干细胞的药物做铺垫。

Objective： To prove that GSCs are widely existed and to suggest a simple strategy for isolating them. Method： Tumor cells were seeded in proper density （100-200 cells） in each well of 96-well plates. We harvested tumorspheres after 2 weeks. Their capability to self-renew was detected. We further determine whether tumorspheres expressed multiple genes enriched in neural and other stem t cells and whether they had the capability to differentiate into multiple lineages by laser scanning microscopy. At last, tumprigenicity of tumorspheres was tested. Results： We found that a small proportion of the cells from several human glioma cell lines could form tumorspheres in culture in a proper density in 96-well plates. These spheres can self-renew and express many genes characteristic of glioma stem cells. Besides, under conditions promoting differentiation, cells are multipotent. In nude mice brains, merely 5,000 cells from tumorspheres could form tumors. Conclusion： Our results further convinced that GSCs are widely existed and will facilitate future studies of initiation and progression of human gliomas.