摘要
目的从人脑肿瘤组织中分离培养脑肿瘤干细胞(braintumor stemcells,BTSCs),对其进行鉴定,并对BTSCs的生物学特性进行探讨。方法取新鲜人脑胶质母细胞瘤标本,分离脑肿瘤细胞,接种于含生长因子的无血清培养基中培养,神经球形成率分析神经球的自我更新及增殖能力;免疫荧光方法检测神经球CD133、Nestin表达情况;神经球细胞经诱导分化后细胞表面分化标记物β-TubulinⅢ、GFAP表达情况。结果在无血清培养基中大部分脑肿瘤细胞死亡,神经球形成率分析只有不到5%细胞具有形成神经球的能力,免疫荧光方法检测神经球CD133、Nestin表达阳性,神经球细胞经诱导分化后β-TubulinⅢ、GFAP表达阳性。结论应用直接培养法获得BTSCs方法简便、易行,可以用于获取BTSCs。
Objective To certify BTSCs and study the biological characteristics of BTSCs,the cells was isolated cultured from fresh brain tumor and cultured with serum-free media.Methods Picked up and dissociated the fresh glioblastoma sample,planted cells in serum-free media with grows factor.The neurosphere forming essay analyzed the self-renewal and proliferative capacity.Immunofluorescent staining detected neural stem cells marker CD133 and Nestin,multipotentiality markers β-TubulinⅢ and GFAP.Results Only about 5% glioblastoma formed the neuroshpere,has the self-renewal and proliferated capacity.Neurosphere was positive when stained with CD133 and Nestin.After differentiation,β-TubulinⅢ and GFAP was expressed.Conclusion Directly isolated and cultured the BTSCs from fresh glioblastoma sample,it is simple and easy,it can be used to obtain BTSCs.
出处
《中国实验诊断学》
北大核心
2011年第2期227-228,共2页
Chinese Journal of Laboratory Diagnosis
基金
吉林省自然科学基金面上项目(202015242)
