摘要
目的探讨时间因素对体外培养的神经干细胞向γ氨基丁酸(gammaaminobutyricacid,GABA)能神经元分化的影响。方法体外培养神经干细胞并加以鉴定。利用反义寡核苷酸诱导神经干细胞向GABA能神经元分化,通过免疫荧光染色计算不同时间点的GAD65阳性细胞率。结果体外培养获得神经干细胞,染色可见神经球及分化细胞中分别有nestin、NF200、GFAP阳性细胞。方差分析表明不同时间点的阳性细胞率存在的差别具有显著意义(F=702.93,P<0.05),LSDt检验表明除第7d与第9d无明显差别外,其余各时间点之间均有明显差别。其中第7天的分化率最高为87.60%±6.26%。结论本实验利用反义寡核苷酸序列特异性阻断了Hes1的表达,促进了神经干细胞向GABA神经元分化,并发现在诱导分化7d后分化率达到最高值。
Objective To investigate the effect of time factor on differentiation of neural stem cells（NSCs） to gamma aminobutyric acid （GABA） neurons. Methods Cultured NSCs in vitro and identified it by immunoflurescence stain. Induced NSCs to differentiate to GABA neurons with antisense oligonucleotides. Then counted the GAD65 positive cell differentiation rate in different time point through immunoflurescence stain. Results Attained NSCs through cultured in vitro. Immunofluorescence stain showed nestin,NF200, GFAP positive cells in nerve bulb or differentiated cells. ANOVA showed that there was significant difference between the positive cell rate of different time points（F =702.93, P 〈 0.05）. LSD-t showed that there was significant difference between every time point except between day 7 and day 9. The highest rate was found in day 7, which was 87.60 % ＋ 6.26 %.Conclusion Antisense oligonucleotides can specifically block Hesl, which promote the differentiation from NSCs to GABA neurons. And we found the positive cell rate climaxed when the cell induced by 7days.
出处
《立体定向和功能性神经外科杂志》
2005年第4期212-215,共4页
Chinese Journal of Stereotactic and Functional Neurosurgery
基金
西京医院临床高新技术面上项目(编号:XJGX03033M25)
关键词
神经干细胞
分化
GABA
神经元
时间因素
Neural stem cells
Differentiation
Gamma aminobutyric acid
Neurons
Time factor
