神经干细胞诱导分化过程中巢蛋白表达的时程变化(英文)

Temporary expression of nestin during neural stem cell differentiation

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:Recently, many studies are focused on the dynamic changes of nestin expression when neural stem cells(NSC) and neural progenitor cells(NPC) differentiate into neurons and gliacytes. OBJECTIVE:To investigate the dynamic changes of nestine expression during NSC induced differentiation in vitro. DESIGN:Repeated measurements. SETTINGS and MATERIALS:The study was performed in Beijing Neurosurgery Institute. Wistar rats(male or female) with body mass around(180±20) g were used for the study.Expression level of nestin at day 3 and at week 1,2,3,4,5,6,7 and 8 had been measured after fixation of cultured cells.Data at each time point were collected from 4 culture wells with 3 repeated measurements. INTERVENTIONS:Cell morphology was recorded between 3 days and 8 weeks after plasma induction.Nestin expression was measured by using immunofluorescence staining. MAIN OUTCOME MEASURES:Level of nestin at different time points of NSC differentiation. RESULTS:Nestin expression was positive during early stage of NSC differentiation.Nestin expression was decreased gradually with the differentiation and increased maturation of neurocytes,astrocytes and oligodendrocytes. Nestine level was deceased to 0 at week 7 after plasma induction. CONCLUSION:Nestin expression is an important marker of NSC differentiation.In addition,nestin is also expressed during the early stage of NSC differentiation,and the expression level decreases with the increased maturation of neurocytes.The level of nestin is down to 0 at week 7 of cell culture.