小鼠胚胎神经干细胞的分离培养及其鉴定

且的探索小鼠胚胎神经干细胞的体外培养方法，并获取高纯度的神经干细胞，为神经干细胞的深入研究提供实验材料。方法无菌条件下分离E15天小鼠胚脑皮质，制成单细胞悬液，在bFGF和B27存在的培养基中培养扩增，通过免疫细胞化学染色鉴定神经干细胞及其子代细胞的分化方向。结果培养的部分细胞在B27和bFGF存在的无血清培养基中可以在体外分裂增殖，同时表达神经干细胞特异性抗原nestin，并在撤出B27和bFGF的有血清培养基中向神经细胞和胶质细胞分化。结论小鼠胚脑皮质存在具有多向分化潜能的神经干细胞，这些细胞可以在体外稳定培养、传代并自然分化，为细胞替代治疗提供了理想的细胞来源。

ISOLATION CULTIVATION AND IDENTIFICATION OF NEURAL STEM CELLS FROM EMBRYONIC MICE

Objective To explore the culture methods of neural stem cells from embryonic mice and to obtain highly purified neural stem cells for further investigation. Method Embryonic corticesof mice were dissociated and single cell suspensions were achieved in sterile conditions. Then the cells were cultured in DMEM/F12 supplemented with bFGF and B27. Immunocytochemical techniques were used to identify stem cells and their progeny. Result In the serum-free medium containing bFGF and B27, the cells from embryonic mice divided and expanded in vitro, in which nestin antigen was expressed. In the containing serum medium but neither bFGF nor B27, the cells differentiated into neurons and glia. Conclusion Neural stem cell from the cortices of embryonic mice can be cultured, passaged and differentiated in vitro. They are the ideal cell sources for cell transplantation therapy.