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(P<0.05)。
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。
Objective
To explore whether the neural stem cells (NSCs) could survive in artificial cerebrospinal fluid (ACSF).
Methods
After in vitro expansion for two passages in standard media, NSCs were respectively seeded and cultured in the ACSF or normal neural media. The characteristics of NSCs growing in the two media were observed and compared. Immunofluorescence staining was adopted to detect the expression of Tubulin and GFAP of cells in two media at the third day.
Results
The NSCs survived and grew in ACFS. The immunofluorescence staining showed Tubulin(+) and GFAP(+). But NSCs in ACFS had a slender body and synaptic compared with those in ordinary neural media.
Conclusion
In this experiment condition, the NSCs could survive, grow and differentiate in ACFS.
Objective
To explore the optimal time point for neural stem cells (NSCs) to transplant in artificial cerebrospinal fluid (ASCF) in vitro.
Methods
After in vitro expansion for two passages in standard media, NSCs were induced to differentiate, and then they were harvested respectively from day 1 to day 6 in the induced differentiation. Cells were seeded in the ASCF and cultured for another 3 days. The positive rate of neuron from the NSCs was calculated. MTT was adopted to explore the relative growth activity of the cells.
Results
In this experiment condition NSCs survived and grew in the ASCF. Statistical analysis showed cells differentiated for 4 days got a significantly higher positive rate of neuron and relative growth activity in ASCF than other time points (P<0.05).
Conclusion
In this experiment condition differentiation for 4 days was the optimal time point for NSCs to transplant in ASCF in vitro
引文
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