《中国康复理论与实践》

《中国康复理论与实践》 ›› 2015, Vol. 21 ›› Issue (04): 406-411.

• 基础研究 • 上一篇    下一篇

碱性成纤维细胞生长因子-壳聚糖载体诱导神经干细胞向神经元分化并形成突触的研究

王聪 1，杨朝阳 1，段红梅 2，李晓光 1   

  1. 1.首都医科大学神经生物学系，北京市 100069；2.北京航空航天大学生物医学工程学院，北京市 100191。
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2015-04-25 发布日期:2015-04-25

Basic Fibroblast Growth Factor-Chitosan Carriers Induce Neural Stem Cells to Differentiate into Neurons and Form Synapses

WANG Cong1, YANG Zhao-yang1, DUAN Hong-mei2, LI Xiao-guang1
   

  1. 1. Department of Neurobiology, Capital Medical University, Beijing 100069, China; 2. Department of Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, China
  • Received:1900-01-01 Revised:1900-01-01 Published:2015-04-25 Online:2015-04-25

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被引次数

7

摘要： 目的 探讨碱性成纤维细胞生长因子(bFGF)-壳聚糖载体对神经干细胞(NSCs)的诱导分化作用。方法 原代培养新生24 h Wistar大鼠脊髓 NSCs，纯化后分别向培养基加入单纯壳聚糖、单纯 bFGF和 bFGF-壳聚糖载体。3 d后，行 Nestin和β-微管蛋白Ⅲ免疫荧光染色；7 d后，行微管相关蛋白2 (MAP2)、神经胶质纤维酸性蛋白(GFAP)和髓鞘碱性蛋白(MBP) 免疫荧光染色；14 d后，行 synapsin-1和 MAP2免疫荧光染色，并利用 MED64平面微电极阵列记录系统检测分化后神经元的电生理活性。结果 培养3 d 后，各组均可见 Nestin+/β-微管蛋白Ⅲ+细胞，bFGF-壳聚糖载体诱导的细胞神经丝长度大于其他两组；7 d 后，各组均可见MAP2+、GFAP+和 MBP+细胞，bFGF-壳聚糖载体诱导的细胞 MAP2+比例高于其他两组；14 d后，bFGF-壳聚糖载体诱导的细胞呈synapsin-1+/MAP2+，且有自发放电现象。结论 bFGF-壳聚糖载体诱导NSCs高比例向神经元分化，且分化而成的神经元之间形成突触，并具有电生理活性。

关键词: 神经干细胞, 碱性成纤维细胞生长因子, 壳聚糖, 神经分化, 突触, 神经元

Abstract: Objective To explore the effect of basic fibroblast growth factor (bFGF)-chitosan carriers on neural differentiation of neural stem cells (NSCs). Methods NSCs were isolated from spinal cord of a neonatal Wistar rat and cultured. Purity of cultured NSCs was identified with Nestin immunofluorescent staining. The 10 mg/ml chitosan carriers, 20 ng/ml bFGF or 10 mg/ml bFGF-chitosan carriers were added into medium of P3~P4 NSCs respectively. NSCs were observed with immunofluorescent staining: 3 days after incubation with Nestin and β-tubulin III; 7 days after incubation with microtubule-associated protein-2 (MAP2), glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP); and 14 days after incubation with synapsin-1 and MAP2. The electrophysiological activity of cells was detected with MED64. Results 3 days after incubation, all the NSCs differentiated into Nestin+/β-tubulin III+, and the length of neurofilament was the highest in those co-cultured with bFGF-chitosan carriers. 7 days after incubation, NSCs differentiated into MAP2+, GFAP+ and MBP+, and more NSCs differentiated into MAP2+ with bFGF-chitosan carriers. 14 days after incubation, NSCs differentiated with bFGF-chitosan carriers express synapsin-1+/MAP2+ and showed electrophysiological activity. Conclusion bFGF-chitosan carriers can induce NSCs to differentiate into neuron with high percentage and the differentiated neurons can form synapses with electrophysiology activity.

Key words: neural stem cells, basic fibroblast growth factor, chitosan, neural differentiation, synapses, neurons