Differential development of neuronal physiological responsiveness in two human neural stem cell lines.
Differential development of neuronal physiological responsiveness in two human neural stem cell lines.
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在两个人类神经干细胞系中神经元生理反应性的差异发展。
DOI：
10.1186/1471-2202-8-36
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发表时间：
2007-05-25
期刊：
影响因子：
2.4
通讯作者：
Sinden, John D.
中科院分区：
文献类型：
--
作者：
Donato, Roberta;Miljan, Erik A.;Hines, Susan J.;Aouabdi, Sihem;Pollock, Kenneth;Patel, Sara;Edwards, Frances A.;Sinden, John D.
神经干细胞（NSCs）是设计和发现神经退行性疾病新方法的有力研究工具。myc家族转录因子在人原代细胞中的过表达，从发展中的皮层和中脑产生了两个稳定的多潜能的神经干细胞系（ReNcell VM和CX），可以在单层培养中不断扩大。在未分化状态下，ReNcell VM和CX都是巢蛋白阳性的，并且具有约-60 mV的静息膜电位，但不显示任何电压激活的电导。正如最初假设的那样，使用标准方法（stdD）进行分化，根据免疫组织学特征，两种细胞系均可形成神经元、星形胶质细胞和少突胶质细胞。然而，很明显，这对指定神经元的电生理特征（如动作电位的放电）来说是不正确的。因此，我们已经开发了一种新的分化方案，命名为“预聚集分化”（preD），这似乎有利于电生理功能神经元的发展，并导致多巴胺能神经元的ReNcell VM线的增加。相反，所用的方案对ReNcell CX的分化几乎没有影响，其中未观察到多巴胺能分化。此外，在用preD方案分化一周后，100%的ReNcell VM具有TTX敏感的Na+通道和激发的动作电位，而stdD后为25%。通过其他电压门控通道的电流似乎不依赖于分化协议。ReNcell CX没有显示出与VM线相同的电生理特性，在stdD或preD分化下产生电压依赖性K+电流，但没有Na+电流或动作电位。这些数据表明，myc在NSC中的过表达可用于在培养物中产生电生理活性神经元。功能性神经元表型的发展可能依赖于细胞系的分离和分化参数，这表明并非所有的人NSC在功能上都是等同的。
Neural stem cells (NSCs) are powerful research tools for the design and discovery of new approaches to neurodegenerative disease. Overexpression of the myc family transcription factors in human primary cells from developing cortex and mesencephalon has produced two stable multipotential NSC lines (ReNcell VM and CX) that can be continuously expanded in monolayer culture. In the undifferentiated state, both ReNcell VM and CX are nestin positive and have resting membrane potentials of around -60 mV but do not display any voltage-activated conductances. As initially hypothesized, using standard methods (stdD) for differentiation, both cell lines can form neurons, astrocytes and oligodendrocytes according to immunohistological characteristics. However it became clear that this was not true for electrophysiological features which designate neurons, such as the firing of action potentials. We have thus developed a new differentiation protocol, designated 'pre-aggregation differentiation' (preD) which appears to favor development of electrophysiologically functional neurons and to lead to an increase in dopaminergic neurons in the ReNcell VM line. In contrast, the protocol used had little effect on the differentiation of ReNcell CX in which dopaminergic differentiation was not observed. Moreover, after a week of differentiation with the preD protocol, 100% of ReNcell VM featured TTX-sensitive Na+-channels and fired action potentials, compared to 25% after stdD. Currents via other voltage-gated channels did not appear to depend on the differentiation protocol. ReNcell CX did not display the same electrophysiological properties as the VM line, generating voltage-dependant K+ currents but no Na+ currents or action potentials under either stdD or preD differentiation. These data demonstrate that overexpression of myc in NSCs can be used to generate electrophysiologically active neurons in culture. Development of a functional neuronal phenotype may be dependent on parameters of isolation and differentiation of the cell lines, indicating that not all human NSCs are functionally equivalent.
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影响因子：
5.3
作者：
Shihabuddin, LS;Ray, J;Gage, FH
通讯作者：
Gage, FH
DOI：
10.1073/pnas.0404700101
发表时间：
2004-08-24
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
2.7
作者：
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通讯作者：
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影响因子：
2.9
作者：
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通讯作者：
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影响因子：
3.3
作者：
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通讯作者：
Fine, A