目的 :建立一个稳定的神经干细胞系 .方法 :从孕12～ 16d小鼠胚胎脑内分离纹状体 ,吹打消化为单细胞悬液 ,于DMEM/F12 (1∶1)培养基 (含有bFGF ,EGF和N2添加剂 )中培养 .通过MTT法检测细胞活性 ,并绘制生长曲线 .采用免疫荧光法观察神经干细胞nestin的表达情况和其分化的子代细胞NSE及GFAP等的表达情况 .结果 :原代分离培养 4～ 6d后 ,培养基内可见神经克隆球 ,分散克隆球所获的单细胞继续培养可以重新生成克隆球 .新生克隆球细胞呈nestin阳性 ,传代入含血清的培养基后可以分化为NSE和GFAP阳性细胞 .结论 :所获新生克隆球细胞是神经干细胞 OBJECTIVE: To establish a stable neural stem cell line.Methods: The striatum was isolated from the embryonic brain of 12 ~ 16 d pregnant pregnant women, and the cells were detached into single cell suspension in DMEM / F12 (1: 1) medium containing bFGF, EGF and N2), and the cell viability was measured by MTT assay, and the growth curve was drawn.Expression of nestin in neural stem cells and the expression of NSE and GFAP in differentiated progeny cells were observed by immunofluorescence.Results: After isolated and cultured for 4 ~ 6 days, the neuronal cloning spheres were observed in the culture medium, and the single cells obtained by dispersing the spheres were regenerated to regenerate the cloned spheres. The newborn clonal spheroids were nestin positive and subcultured into serum-containing medium and then differentiated into NSE and GFAP positive cells.Conclusion: The newborn clonal sphere cells are neural stem cells