Plasticity of Calcium Signaling Cascades in Human Embryonic Stem Cell-Derived Neural Precursors
Plasticity of Calcium Signaling Cascades in Human Embryonic Stem Cell-Derived Neural Precursors
复制标题
DOI：
10.1089/scd.2012.0624
复制
发表时间：
2013-05-01
影响因子：
4
通讯作者：
Dayanithi, Govindan
中科院分区：
文献类型：
--
作者：
Forostyak, Oksana;Romanyuk, Nataliya;Dayanithi, Govindan
人胚胎干细胞来源的神经前体细胞（hESC NPs）被认为是基于细胞的治疗中枢神经系统损伤和神经退行性疾病的有希望的工具。Ca 2+离子是调节各种细胞功能所必需的重要细胞内信使。我们研究了Ca 2+信号传导的作用和生理学，以表征在培养（体外）中长期维持期间CCTL 14 hESC NP的功能特性。我们分析了在hESC分化过程中由高K+、腺苷-5 '-三磷酸（ATP）、谷氨酸、γ-氨基丁酸（GABA）和咖啡因诱发的细胞质Ca 2+浓度（[Ca 2 +]（i））的变化与不同代次（P6至P10）中各种神经元标记物表达的相关性。我们发现只有来自P7的分化的NPs对各种刺激表现出显著和特异的[Ca 2 +]（i）反应。约31%的神经元样P7 NPs表现出自发的[Ca 2 +]（i）振荡。药理学和免疫细胞化学分析显示，P7 NPs表达L型和P/Q型Ca 2+通道、P2 X（2）、P2 X（3）、P2 X（7）和P2 Y嘌呤受体、谷氨酸受体和兰尼碱（RyR 1和RyR 3）受体。ATP和谷氨酸诱导的[Ca 2 +]（i）反应具有浓度依赖性。较高的谷氨酸浓度（超过100 μ M）引起细胞死亡。在存在或不存在细胞外Ca 2+的情况下观察对ATP的反应。这些结果强调了这样的概念，即随着培养时间的推移，这些细胞获得了短暂的Ca 2+信号传导，这预示着它们作为干细胞的能力。
Human embryonic stem cell-derived neural precursors (hESC NPs) are considered to be a promising tool for cell-based therapy in central nervous system injuries and neurodegenerative diseases. The Ca2+ ion is an important intracellular messenger essential for the regulation of various cellular functions. We investigated the role and physiology of Ca2+ signaling to characterize the functional properties of CCTL14 hESC NPs during long-term maintenance in culture (in vitro). We analyzed changes in cytoplasmic Ca2+ concentration ([Ca2+](i)) evoked by high K+, adenosine-5'-triphosphate (ATP), glutamate, gamma-aminobutyric acid (GABA), and caffeine in correlation with the expression of various neuronal markers in different passages (P6 through P10) during the course of hESC differentiation. We found that only differentiated NPs from P7 exhibited significant and specific [Ca2+](i) responses to various stimuli. About 31% of neuronal-like P7 NPs exhibited spontaneous [Ca2+](i) oscillations. Pharmacological and immunocytochemical assays revealed that P7 NPs express L-and P/Q-type Ca2+ channels, P2X(2), P2X(3), P2X(7), and P2Y purinoreceptors, glutamate receptors, and ryanodine (RyR1 and RyR3) receptors. The ATP- and glutamate-induced [Ca2+](i) responses were concentration-dependent. Higher glutamate concentrations (over 100 mu M) caused cell death. Responses to ATP were observed in the presence or in the absence of extracellular Ca2+. These results emphasize the notion that with time in culture, these cells attain a transient period of operative Ca2+ signaling that is predictive of their ability to act as stem elements.