小鼠胚胎神经干细胞的长期培养和分化

目的：探索小鼠胚胎神经干细胞的体外培养和分化条件,为神经干细胞的深入研究奠定基础。方法：无菌条件下分离小鼠胚胎脑皮质,制成单细胞悬液,种植在N2培养基中培养,培养基中加入20ng/ml的碱性成纤维细胞生长因子（每次实验用1－2只小鼠胚胎,实验重复8次）。采用机械方法传代,常规方法冻存和复苏。用免疫细胞化学方法对培养的细胞进行鉴定。结果：成功培养出小鼠的神经干细胞,神经干细胞呈悬浮状态生长,形成典型的神经球。细胞表达巢蛋白和波形蛋白2种神经干细胞的标志物。细胞在胎牛血清的诱导下,可分化成神经元、星型胶质细胞和少突胶质细胞,它们所占的比例分别为7％、85％－90％和2％－4％。结论:鼠的神经干细胞可以在体外稳定地培养和传代,为细胞治疗提供了一个理想的细胞来源。

Long-term culture and differentiation of neural stem cells of embryonic mice

Objectives To assess the culture and differentiation of neural stem cells in embryonic mice and set up a basis for further reseach in to neural stem cells. Methods Embryonic cortices of mice were dissociated and single cell suspensions were achieved by mechanical methods in sterile conditions, and cells were seeded in uncoated plate in N2 medium. The cells were passaged by mechanical methods, frozen and thawed by general procedure. They were identified by immunocytochemical techniques. Results Neural stem cells from embryonic mice were successfully cultured forming typical neurospheres in suspension. Neurons, astrocytes and oligodendrocytes were differentiated from neural stem cells, with a ratio of 7%, 85% 90% and 2% 4% respectively. Conclusions Neural stem cells, which can be cultured and passaged steadily in vitro and they are the ideal cell sources for cell transplantation and gene therapy.