胶质瘤干细胞的培养和分化研究

目的探讨人类胶质瘤干细胞的体外培养、传代和分化。方法采用机械方法从胶质瘤组织中分离细胞，应用N2培养基进行培养，碱性成纤维细胞生长因子和表皮生长因子刺激细胞扩增；应用免疫组织化学染色对培养的细胞及其分化的细胞进行鉴定。结果从胶质瘤组织当中成功培养出人类的胶质瘤干细胞，培养条件下呈悬浮状态生长，形成神经球，绝大多数的细胞表达波形蛋白和巢蛋白两种神经干细胞的标志物；这种细胞可分化为神经元和星型胶质细胞。结论体外的培养条件下，可从胶质瘤组织中培养出干细胞，能够分化为神经元和星型胶质细胞，为胶质瘤的深入研究莫定基础。

Long-term Culture and Differentiation of Human Glioma Stem Cells

OBJECTIVE: To explore the culture and subculture conditions for glioma stem cells(GSCs) and to investigate the differentiation potential of GSCs. METHODS: The cells from human glioma were mechanically dissociated. Cells were cultured in N2 or B27 medium with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF), and they were identified by immunocytochemistry. RESULTS: Glioma stem cells from human glioma have been successfully cultured. They formed typical neurospheres in suspension, and they could be cultured and passaged steadily in vitro. The majorities of the cells expressed vimentin and nestin, which were the markers for neural stem cells. They could differentiate into neurons and astrocytes, and express glial fibrillary acidic protein and beta III-tubulin respectively. CONCLUSION: Human glioma stem cells could be cultured from gliomas in vitro, and they could differentiate into neurons and astrocytes, thus providing a basis for further studies.