成年大鼠嗅球神经干细胞的培养和鉴定

Culture and Identification of Neural Stem Cell Derived from Olfactory Bulb in Adult Rat

目的建立成年大鼠嗅球神经干细胞分离培养和鉴定方法,探索新的成年神经干细胞种子来源。方法用无血清方法分离培养成年大鼠嗅球来源的神经干细胞;用克隆培养、BrdU整合的方法检验培养细胞的干细胞特性;用免疫荧光细胞化学的方法检测BrdU、神经干细胞标记物Nestin和SOX2,分化的细胞标记物Tuj1、GFAP、NG2的表达。结果从成年大鼠嗅球能够分离、培养出具有自我更新、增殖的神经球,构成神经球的细胞呈Nestin和SOX2阳性,它们分化后产生Tuj1阳性的神经元、GFAP阳性的星形胶质细胞、NG2阳性的少突胶质细胞。结论成年大鼠嗅球存在神经干细胞,能够在体外进行培养、增殖、分化,是神经干细胞的新的种子来源。

Objective To establish a method to culture neural stem cells（NSCs） derived from olfactory bulb（OB） in adult rat and identify the property of NSCs.Methods NSCs were isolated from OB of adult rat and cultured in serum-free medium.Clone culture and BrdU incorpoion were performed to identify the self-renewal and proliferation of NSCs.Fluorescence immunocytochemistry was carried out to examine the expression of NSCs marker nestin,SOX2,neuronal marker Tuj1,astrocyte marker GFAP and oligodendrocyte marker NG2.Results Self-renewal and prolifered NSCs were obtained from OB in adult rat and grown as floating neurospheres.They expressed neural stem cell marker nestin and SOX2 and can differentiated to Tuj1 positive neurons,GFAP positive astrocytes and NG2 positive oligodendrocytes.Conclusion NSCs exist in OB of adult rat and they showed stem cell property.