低氧联合GDNF诱导大鼠神经干细胞分化为TH阳性神经元的实验研究  

Combined use of low oxygen and GDNF on induction of differentiation from neural stem cells into dopaminergic neurons in vitro

作　　者：杜娟[1] 刘昌红[2] 梁辉[1] 吕晓红[3] DU Juan;LIU Changhong;LIANG Hui(Yantaishan Hospital of Shandong,Yantai 264008,China)

机构地区：[1]山东省烟台市烟台山医院神经内科,山东烟台264008 [2]山东省烟台市烟台山医院物资材料供应科,山东烟台264008 [3]吉林大学白求恩第一医院神经内科,吉林长春130021

出　　处：《中风与神经疾病杂志》2021年第4期322-325,共4页Journal of Apoplexy and Nervous Diseases

基　　金：烟台市科技创新计划(No.2015YT06000365)。

摘　　要：目的本试验在低氧环境下利用胶质源性神经营养因子(GDNF)对大鼠胚胎神经干细胞(NSCs)进行定向诱导分化,从而研究低氧对NSCs向多巴胺能神经元分化的影响,并获得相当数量的多巴胺能细胞,为下一步的细胞移植奠定基础。方法分离培养孕14 d~16 d Wistar大鼠胚胎皮质NSCs,通过传代培养获得单克隆生长的NSCs,在不同氧浓度下利用GDNF对大鼠NSCs进行定向诱导分化,通过免疫细胞化学方法检测TH阳性细胞表达。结果低氧可明显促进NSCs向TH阳性神经元分化,与常氧对照组比较有显著差异(P<0.01),且在3%低氧浓度时,NSCs向TH阳性神经元分化率最高。结论低氧可促进NSCs向TH阳性神经元分化,3%低氧浓度是促进NSCs向TH阳性神经元分化的最适氧浓度,低氧促进NSCs向TH阳性神经元分化的分化率与氧浓度梯度无依赖关系,低氧联合GDNF诱导NSCs分化为TH阳性神经元,诱导效能更为显著。Objective In this study,glial cell-derived neurotrophic factor(GDNF)were used to induce neural stem cells(NSCs)derived from brain tissue of rat embryo to directional differentiation at a low oxygen environment.Thereby,exploring the affects of low oxygen on differentiation of NSCs into dopaminergic neurons.In this condition.We got quite a lot of dopaminergic cells,which will lay the foundation for the next step of cell transplantation.Methods NSCs were isolated and cultured from cortex of 14~16 days Wistar rat embryo.Then,at a series of different oxygen concentration with adding of GDNF,the TH positive cells differentiated from NSCs were detected by immunocytochemistry and immunofluorescence methods.Results Low oxygen obviously increased TH expression with significant differences compared to the normal oxygen control group(P<0.01).What’s more,at 3%low oxygen concentration,there was a highest differentiation rate for NSCs to TH positive neurons.Conclusion Low oxygen concentration promoted the differentiation from NSCs to TH positive neurons.3%oxygen concentration was the optimum low oxygen concentration.There was no dependence relationship between differentiation rate promoted by low oxygen concentration for NSCs to TH positive neurons and oxygen concentration gradient.There was a more significant increase in induction efficiency for combined use of low oxygen and GDNF on induction of differentiation of NSCs into TH positive neurons.

关 键 词：神经干细胞 低氧 多巴胺能神经元 定向分化 

分 类 号：R741[医药卫生—神经病学与精神病学]