白细胞介素-2对神经干细胞增殖的影响

发布时间：2018-02-11 02:08

本文关键词： 神经干细胞白细胞介素-2 增殖　出处：《浙江大学》2010年硕士论文　论文类型：学位论文

【摘要】：目的提取胚胎大鼠(E14)大鼠脑室下区、海马等部位的神经干细胞体外条件下进行原代培养和传代培养,并研究白细胞介素-2(interleukin-2, IL-2)对神经干细胞增殖的影响。方法提取胚胎大鼠(E14)大鼠脑室下区、海马等部位的神经干细胞,体外条件下在完全培养基中进行原代培养和传代培养,培养后行nestin染色,检测其是否为神经干细胞。将细胞分为四组,实验组内分别添加一定量的IL-2使其浓度分别为50ng/ml、100ng/ml和150ng/ml;对照组不添加IL-2。相同条件下培养3天后,加入BrdU,终浓度10μmol/L;继续培养1天后行免疫细胞化学染色；显微镜下观察,并计数阳性细胞和阴性细胞数,计算各组神经干细胞的增殖率。结果原代培养的神经干细胞数目较少,培养第一天就可见大量细胞贴壁,少数细胞伸出突起,从培养的第4-5天开始,贴壁的细胞破裂、死亡,而未贴壁的细胞则悬浮于培养基中,细胞数目增多,形成小的神经球(Neurosphere),神经球逐渐增大。传代细胞呈球形悬浮于培养瓶中,细胞形态规则,未见明显突起生长,传代7天时神经球已生长成为数百个细胞的大克隆。神经球nestin免疫细胞化学染色基本全部呈阳性,证实其为神经干细胞。加入IL-2后神经干细胞的增殖率显著增加,其中IL-2浓度为150ng/ml时,神经干细胞的增殖率最大,明显高于对照组；IL-2浓度为50ng/ml时神经干细胞的增殖率较IL-2浓度为150ng/ml时低,但仍显著高于对照组；IL-2浓度为100ng/ml时神经干细胞的增殖率介于前两组之间,且显著高于对照组。结论一定浓度的IL-2可以促进神经干细胞的增殖。
[Abstract]:Objective to extract neural stem cells (NSCs) from the subventricular and hippocampal regions of embryonic rat (E14) in vitro and to study the effects of interleukin-2 (IL-2) on the proliferation of neural stem cells (NSCs). Methods the neural stem cells were extracted from the subventricular area and hippocampus of the embryonic rat (E14). The neural stem cells were cultured and subcultured in a complete culture medium in vitro. Nestin staining was performed after culture. The cells were divided into four groups. In the experimental group, a certain amount of IL-2 was added to the cells at the concentrations of 50 ng / ml 100 ng / ml and 150 ng / ml, respectively, while the control group was not supplemented with IL-2.After the same conditions, the cells were cultured for 3 days. BrdU was added at the final concentration of 10 渭 mol / L; immunocytochemical staining was performed after 1 day of culture; the number of positive and negative cells was observed under microscope, and the proliferation rate of neural stem cells in each group was calculated. Results the number of primary cultured neural stem cells was relatively small. On the first day of culture, a large number of cells adhered to the wall and a few cells protruded. From the 4-5 days after culture, the adherent cells broke down and died. However, the unattached cells were suspended in the culture medium, the number of cells increased, and the small neurosphere was formed, and the neurosphere gradually increased. The passage cells were spherical suspension in the culture flask, the morphology of the cells was regular, and there was no obvious protruding growth. After 7 days of passage, the nerve ball had grown into a large clone of hundreds of cells. The immunocytochemical staining of nestin was positive, which confirmed that it was a neural stem cell. The proliferation rate of neural stem cells increased significantly after the addition of IL-2. When the concentration of IL-2 was 150ng / ml, the proliferation rate of neural stem cells was the highest, which was significantly higher than that of the control group when the concentration of IL-2 was 50ng / ml, and the proliferation rate of neural stem cells was lower than that of IL-2 (150ng / ml). But the proliferation rate of neural stem cells was significantly higher than that of the control group when the concentration of IL-2 was 100 ng / ml, and the proliferation rate of neural stem cells was significantly higher than that of the control group. Conclusion certain concentration of IL-2 can promote the proliferation of neural stem cells.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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