摘要
目的 体外分离、培养和鉴定胎小鼠神经干细胞(NSC)并检测其一氧化氮合酶 (NOS)的表达 ,为进一步阐明NOS在NSC中的作用奠定基础 .方法 利用含EGF的条件培养基 ,从胚胎小鼠大脑皮层中分离并体外培养胎小鼠NSC ,在含胎牛血清的培养基中诱导其分化 ;用充纯氮气的方法造成细胞缺氧 ,观察缺氧对NOS的诱导作用 ;免疫组织化学方法检测细胞中nestin ,nNOS ,eNOS和iNOS的表达 .结果 原代培养的细胞具有连续增殖并形成克隆的能力 ,在含血清的培养基中能分化为神经元及神经胶质细胞样细胞 ;免疫组化nestin ,nNOS和eNOS在一般培养条件下呈阳性表达 ,而iNOS呈阴性表达 ,经充氮气缺氧诱导后 ,iNOS呈阳性表达 .结论 成功分离并培养小鼠NSC ;发现不同亚型的NOS在NSC不同培养条件下有不同的表达 .
AIM To isolate, cultivate and identify the neural stem cells(NSC) from the cortex of embryo mouse in vitro, and examine the nitric oxide synthase(NOS) expressing in it , for the further explanation of the effects of NOS expressing in NSC. METHODS The NSC was isolated from the cortex of embryo mouse and grew in mediums contain EGF , and was induced to differentiate in mediums supplemented with fetal bovine serum; The culture flask were inflated with pure nitrogen gas to cause the cells hypoxia, to observe the induced effects of hypoxia on NOS; The expression of nestin,nNOS,eNOS and iNOS in the NSC was examined by immunohistochemistry methods. RESULTS The primary culture cells have the ability to proliferate continuously and to form neurosphere, and the cells can differentiate into neuron and glial like cells in mediums supplemented with fetal bovine serum; The cells are nestin,nNOS and eNOS positive expression which were detected by immunohistochemistry methods at general culture conditions, whereas the expression of iNOS is negative. After induced by hypoxia, the expression of iNOS is positive too. CONCLUSION We have isolated and cultured the NSC from the cortex of embryo mouse successfully in vitro and found that the several kinds of isoforms of NOS had different expression at various culture conditions.
出处
《第四军医大学学报》
北大核心
2002年第23期2160-2163,共4页
Journal of the Fourth Military Medical University
