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人类诱导多能干细胞衍生的多巴胺能神经元的自动和手动膜片钳数据。

Automated and manual patch clamp data of human induced pluripotent stem cell-derived dopaminergic neurons.

机构信息

Chair of Biophysics, University of Rostock, Rostock 18057, Germany.

Sophion-Biolin Scientific, Ballerup 2750, Denmark.

出版信息

Sci Data. 2017 Apr 25;4:170056. doi: 10.1038/sdata.2017.56.

Abstract

Human induced pluripotent stem cells can be differentiated into dopaminergic neurons (Dopa.4U). Dopa.4U neurons expressed voltage-gated Na and K channels and showed neuron-like spontaneous electrical activity. In automated patch clamp measurements with suspended Dopa.4U neurons, delayed rectifier K current (delayed K) and rapidly inactivating A-type K current (fast K) were identified. Examination of the fast K current with inhibitors yielded IC values of 0.4 mM (4-aminopyridine) and 0.1 mM (tetraethylammonium). In manual patch clamp measurements with adherent Dopa.4U neurons, fast K current could not be detected, while the delayed K current showed an IC of 2 mM for 4-aminopyridine. The Na channels in adherent and suspended Dopa.4U neurons showed IC values for tetrodotoxin of 27 and 2.9 nM, respectively. GABA-induced currents that could be observed in adherent Dopa.4U neurons could not be detected in suspended cells. Application of current pulses induced action potentials in approx. 70 % of the cells. Our results proved the feasibility of automated electrophysiological characterization of neuronal cells.

摘要

人诱导多能干细胞可分化为多巴胺能神经元 (Dopa.4U)。Dopa.4U 神经元表达电压门控钠和钾通道,并表现出类似神经元的自发性电活动。在悬浮的 Dopa.4U 神经元的自动膜片钳测量中,鉴定出延迟整流钾电流 (delayed K) 和快速失活 A 型钾电流 (fast K)。用抑制剂对 fast K 电流的检测得出 IC 值分别为 0.4 mM(4-氨基吡啶)和 0.1 mM(四乙铵)。在贴壁 Dopa.4U 神经元的手动膜片钳测量中,无法检测到 fast K 电流,而延迟 K 电流在 4-氨基吡啶中显示出 2 mM 的 IC 值。贴壁和悬浮的 Dopa.4U 神经元中的钠通道对河豚毒素的 IC 值分别为 27 和 2.9 nM。在贴壁的 Dopa.4U 神经元中可以观察到的 GABA 诱导电流在悬浮细胞中无法检测到。施加电流脉冲可在大约 70%的细胞中诱导动作电位。我们的结果证明了自动电生理特性分析神经元细胞的可行性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f941/5404656/f7a79070cb20/sdata201756-f1.jpg

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