Ion channels and ionotropic receptors in human embryonic stem cell derived neural progenitors.
Ion channels and ionotropic receptors in human embryonic stem cell derived neural progenitors.
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DOI：
10.1016/j.neuroscience.2011.04.039
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发表时间：
2011-09-29
期刊：
影响因子：
3.3
通讯作者：
Stice, S. L.
中科院分区：
文献类型：
--
作者：
Young, A.;Machacek, D. W.;Dhara, S. K.;MacLeish, P. R.;Benveniste, M.;Dodla, M. C.;Sturkie, C. D.;Stice, S. L.
从人胚胎干细胞分化的人神经前体细胞为研究神经退行性疾病和药物筛选试验提供了潜在的细胞来源。以前，我们证明，人类神经祖细胞可以维持在一个增殖状态，白血病抑制因子和碱性成纤维细胞生长因子的加入。在这里，我们证明，96小时后，去除碱性成纤维细胞生长因子的神经祖细胞培养显着改变和细胞复制停止。去除碱性成纤维细胞生长因子后14天，大多数细胞表达MAP 2和TUJ1，这是表征有丝分裂后神经元表型的标志物以及神经发育标志物Cdh 2和Gbx 2。进行实时PCR以确定离子营养型受体亚基表达谱。分化的神经祖细胞表达谷氨酸能、GABA能、烟碱能、嘌呤能和瞬时受体电位受体的亚单位。此外，还表达了钠和钙通道亚基。在功能上，几乎所有的NP细胞测试下全细胞电压钳表现出延迟整流钾通道电流和一些分化的细胞表现出河豚毒素敏感，电压依赖性钠通道电流。在少数细胞中，也可以通过全细胞电流钳下的电流注入引起动作电位。这些结果表明，从神经祖细胞培养物中去除碱性成纤维细胞生长因子导致有丝分裂后状态，并具有产生可产生动作电位的可兴奋细胞的能力，这是神经元表型的标志性特征。这是第一次报告的一个有效的和简单的手段，产生人神经元细胞的离子型受体测定，并最终为电活性的人神经细胞测定药物发现。
Human neural progenitor cells differentiated from human embryonic stem cells offer a potential cell source for studying neurodegenerative diseases and for drug screening assays. Previously, we demonstrated that human neural progenitors could be maintained in a proliferative state with the addition of leukemia inhibitory factor and basic fibroblast growth factor. Here we demonstrate that 96 hours after removal of basic fibroblast growth factor the neural progenitor cell culture was significantly altered and cell replication halted. 14 days after the removal of basic fibroblast growth factor, most cells expressed MAP2 and TUJ1, markers characterizing a post-mitotic neuronal phenotype as well as neural developmental markers Cdh2 and Gbx2. Real-time PCR was performed to determine the ionotrophic receptor subunit expression profile. Differentiated neural progenitors express subunits of glutamatergic, GABAergic, nicotinic, purinergic and transient receptor potential receptors. In addition, sodium and calcium channel subunits were also expressed. Functionally, virtually all the NP cells tested under whole-cell voltage clamp exhibited delayed rectifier potassium channel currents and some differentiated cells exhibited tetrodotoxin-sensitive, voltage-dependent sodium channel current. Action potentials could also be elicited by current injection under whole-cell current clamp in a minority of cells. These results indicate that removing basic fibroblast growth factor from the neural progenitor cell cultures leads to a post-mitotic state, and has the capability to produce excitable cells that can generate action potentials, a landmark characteristic of a neuronal phenotype. This is the first report of an efficient and simple means of generating human neuronal cells for ionotrophic receptor assays and ultimately for electrically active human neural cell assays for drug discovery.
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影响因子：
3.4
作者：
Harrill, Joshua A.;Freudenrich, Theresa M.;Mundy, William R.
通讯作者：
Mundy, William R.
影响因子：
16.2
作者：
GEIGER, JRP;MELCHER, T;MONYER, H
通讯作者：
MONYER, H
DOI：
10.1073/pnas.91.7.2815
发表时间：
1994-03-29
影响因子：
11.1
作者：
CULIAT, CT;STUBBS, LJ;RINCHIK, EM
通讯作者：
RINCHIK, EM
影响因子：
3.7
作者：
Katayama, M;Mizuta, I;Ishii, K
通讯作者：
Ishii, K
DOI：
10.1038/nrn2463
发表时间：
2008-09
期刊：
Nature reviews. Neuroscience
影响因子：
--
作者：
Hébert JM;Fishell G
通讯作者：
Fishell G