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背景:神经干细胞和前体细胞在分化成神经元和胶质细胞过程中巢蛋白表达的动态变化是神经生物学家研究的焦点之一。目的:探讨巢蛋白(nestin)在体外诱导神经干细胞分化过程中表达的变化规律。设计:重复测量设计。地点和对象:实验地点为北京市神经外科研究所;研究对象为Wistar大鼠,雌雄不限,体质量(180±20)g;血清诱导后第3天,1,2,3,4,5,6,7,8周分别固定细胞检测巢蛋白的表达,每个时间点4个培养皿,实验重复3次。干预:血清诱导后第3天～第8周相差显微镜连续观察细胞形态的变化,免疫荧光染色检测巢蛋白表达的变化。主要观察指标:在神经干细胞体外诱导分化的不同时间点检测巢蛋白表达的变化。结果:在神经干细胞分化早期,巢蛋白表达仍呈阳性。随着分化的神经细胞、星形胶质细胞或少突胶质细胞逐渐发育成熟,巢蛋白表达逐渐减弱,到第7周时巢蛋白表达完全消失。结论:巢蛋白表达不仅是神经干细胞的重要标志,同时也出现在神经干细胞分化的早期,而且随着细胞逐渐分化成熟,其表达显著下降。体外培养情况下至分化的第7周即完全消失。
BACKGROUND: Dynamic changes in nestin expression of neural stem cells and precursor cells during their differentiation into neurons and glia are among the focuses of neurobiologists’ research. Objective: To investigate the expression of nestin during the differentiation of neural stem cells induced in vitro. Design: repeat measurement design. Place and subject: The experimental site was Beijing Institute of Neurosurgery. The subjects were Wistar rats, both male and female, body weight (180 ± 20) g; on the 3rd day after the induction of serum, 1,2,3,4,5 At 6, 7, and 8 weeks, the expression of nestin was detected by fixed cells. Four dishes were prepared at each time point. The experiment was repeated 3 times. Intervention: The changes of cell morphology were observed by phase contrast microscopy from the 3rd to the 8th week after induction of serum. The expression of nestin was detected by immunofluorescence staining. MAIN OUTCOME MEASURES: The changes of nestin expression were detected at different time points when neural stem cells were induced to differentiate in vitro. Results: Nestin expression was still positive in the early stage of neural stem cell differentiation. With the differentiation of neurons, astrocytes or oligodendrocytes gradually mature, nestin expression gradually weakened, by the seventh week the nestin expression completely disappeared. Conclusion: Nestin expression is not only an important marker of neural stem cells, but also appears early in the differentiation of neural stem cells, and its expression is significantly decreased as the cells gradually differentiate and mature. In vitro culture to the first 7 weeks of differentiation that completely disappeared.