成年小鼠嗅球神经干细胞的培养和鉴定

目的 建立完善的成年小鼠嗅球神经千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 分离培养成年小鼠嗅球来源的神经干细胞;用克隆培养、5-溴2-脱氧尿嘧啶核昔(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、神经干细胞标记物巢蛋白(nestin)和SOX2、分化的细胞标记物Tuj1、胶质纤维酸性蛋白(GFAP)、04的表达. 结果 从成年小鼠嗅球能够分离、培养出具有自我更新、增殖能力的神经球.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、04阳性的少突胶质细胞. 结论 成年小鼠嗅球存在神经干细胞,其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.

In vitro culture and identification of neural stem cells derived from the olfactory bulb of adult mice

Objecfive To establish a method for in vitro culture and identification of neural stem cells(NSCs)derived from the olfactory bulb(OB)of adult mice and test the possibility of the OB as a new source of seed cells of adult NSCs. Methads NSCs were isolated from the OB of adult mice and cultured in serum-free medium.Clonal culture and BrdU incorporation assay were performed to assess the self-renewal and proliferative activities of the NSCs.Fluorescence immunocytochemistry was carried out to examine the expression of the NSC markers nestin and SOX2,neuronal marker Tujl,astrocyte marker GFAP and oligodendroeyte marker 04. Results NSCs possessing self-renewal and proliferative capacities were obtained from the OB of adult mice,and the cells grew in the form of floating neurospheres in the medium.The neurospheres consisted of cells were positive for NSC markers nestin and SOX2,which Were able to differentiate into Tuj1-positive neurons,GFAP-positive astrocytes and 04-positive oligodendrocytes. Conclusion NSCs are present in the OB of adult mice,and the NSCs isolated from the OB can proliferate and differentiate in vitro with obvious stem cell properties.suggesting the feasibility of using OB as anew source of adult NSCs.