神经干细胞最佳移植时间的初步探讨

发布时间：2018-03-28 14:38

本文选题：神经干细胞　切入点：人工脑脊液　出处：《华中科技大学》2011年硕士论文

【摘要】：目的探讨体外培养的神经干细胞(neural stem cells,NSCs)能否在人工脑脊液(artificial cerebrospinal fluid, ACSF)中生长,从而为进一步的细胞移植治疗神经系统疾病奠定基础。方法体外培养NSCs在传2代时,分别接种到ACSF中和常规神经元培养液中培养,观察、比较NSCs在ACSF中及常规神经元培养液的生长情况,并于培养第3天时进行Tubulin、GFAP免疫荧光染色。结果在此实验条件下,NSCs在ACSF中可以存活、生长,且Tubulin、GFAP免疫荧光染色均为阳性。但细胞形态较在常规神经元培养液中培养的细胞有差别,在ACSF中分化的细胞突起细长,胞体欠饱和。结论在此实验条件下,NSCs在ACSF中可以存活、分化。目的探讨体外培养的神经干细胞(neural stem cells, NSCs)在人工脑脊液(artificial cerebrospinal fluid, ACSF)中的最佳移植时间,为细胞移植治疗神经系统疾病奠定基础。方法体外培养NSCs在传2代时进行诱导分化,分别在诱导分化期开始第1、2、3、4、5、6天时收获细胞,重新接种于ACSF中再培养3 d,检测NSCs在ACSF中分化为神经元的百分率,MTT试验检测细胞相对生长活力,初步探讨NSCs在ACSF中的最佳移植时间。结果在此实验条件下,NSCs在ACSF中可以存活并且能够生长,经统计学分析显示,经诱导分化4 d的NSCs,接种于ACSF后分化为神经元的百分率及生长活力显著高于其他时间点(P0.05)。结论在此实验条件下,NSCs在体外诱导分化后第4天时,可能是最佳移植时间。
[Abstract]:Purpose. To investigate whether neural stem cells can grow in artificial cerebrospinal fluid (cerebrospinal) in vitro, so as to lay a foundation for further cell transplantation in the treatment of nervous system diseases. Method. In vitro culture of NSCs was inoculated into ACSF and conventional neuronal culture medium respectively at the second generation. The growth of NSCs in ACSF and conventional neuronal culture medium was compared, and the immunofluorescence staining of NSCs was performed on the 3rd day of culture. Results. Under these experimental conditions, ACSF could survive and grow, and tubal tubulinine GFAP immunofluorescence staining was positive, but the cell morphology was different from that in normal neuron culture medium. The cells differentiated in ACSF had slender processes and undersaturated cell bodies. Conclusion. Under these experimental conditions, NSCs can survive and differentiate in ACSF. Purpose. To investigate the optimal transplantation time of neural stem cells (NSCs) in artificial cerebrospinal fluid (cerebrospinal) in vitro, and to lay a foundation for the treatment of nervous system diseases by cell transplantation. Method. NSCs was cultured in vitro to induce differentiation at the second generation, and the cells were harvested at the beginning of the first day of differentiation at the beginning of the first day of differentiation. The percentage of NSCs differentiated into neurons in ACSF was detected by reseeding in ACSF for 3 days. The relative growth activity of cells was detected by MTT assay. The optimal transplantation time of NSCs in ACSF was preliminarily discussed. Results. Under these experimental conditions, NSCs could survive and grow in ACSF. Statistical analysis showed that the percentage and growth activity of NSCs differentiated into neurons after 4 days of differentiation after inoculation with ACSF were significantly higher than those at other time points (P0.05). Conclusion. Under these conditions, NSCs may be the best transplantation time on the 4th day after induction and differentiation in vitro.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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