目的分离培养树鼩海马神经干细胞,并对其进行鉴定。方法应用加有B27添加剂、碱性成纤维细胞生长因子和表皮生长因子的培养液对新生树鼩海马部脑细胞进行体外培养,并进行传代。采用免疫组织化学及免疫荧光法对培养细胞的Nestin抗原进行鉴定。结果从新生树鼩海马部分离的细胞可连续传代和不断增殖形成神经球,神经球在含血清培养基中具有多向分化能力,Nestin表达阳性。结论采用特定的培养液可以在体外培养树鼩海马神经干细胞。
Objective To investigate the isolation culture and identification of neural stem cells(NSCs) from hippocampus of tree shrew. Methods The solution mixed with B-27 supplement, the basic fibroblast growth factor (bFGF) and epidermal growth factor(EGF) was applied to culture the hippocampal ceils of neonatal tree shrew in vitro, and continuous passage culture was performed. Immunohistochemistry and immunofluorescence methods were used to identify the Nestin antigen of the cells. Results The cells isolated from the tree shrew~ hippocampus had the ability of serial passage and proliferated continuously to become nerve bulbs ,which was with multi-directional differentiation in the serum culture and expressed Nestin positively. Conclusion NSCs from hippocampus of tree shrew can grow in vitro with special culture solution.