Institute for Pharmacology and Clinical Pharmacy, Biochemical-Pharmacological Center Marburg, University of Marburg, Karl-von-Frisch Straße 1, 35032 Marburg, Germany.
J Neurosci Methods. 2012 Jan 15;203(1):69-77. doi: 10.1016/j.jneumeth.2011.09.012. Epub 2011 Sep 22.
Detection of neuronal cell death is a standard requirement in cell culture models of neurodegenerative diseases. Although plenty of viability assays are available for in vitro applications, most of these are endpoint measurements providing only little information on the kinetics of cell death. Here, we validated the xCELLigence system based on impedance measurement for real-time detection of cell death in a neuronal cell line of immortalized hippocampal neurons (HT-22 cells), neuronal progenitor cells (NPC) and differentiated primary cortical neurons. We found a good correlation between impedance measurements and endpoint viability assays in HT-22 cells and NPC, for detecting proliferation, cell death kinetics and also neuroprotective effects of pharmacological inhibitors of apoptosis. In primary neurons we could not detect dendritic outgrowth during differentiation of the cells. Cell death in primary neurons was detectable by the xCELLigence system, however, the changes in the cell index on the basis of impedance measurements depended to a great extent on the severity of the insult. Cell death induced by ionomycin, e.g. shows as a fast paced process involving a strong cellular disintegration, which allows for impedance-based detection. Cell death accompanied by less pronounced morphological changes like glutamate induced cell death, however, is not well accessible by this approach. In conclusion, our data show that impedance measurement is a convenient and reliable method for the detection of proliferation and kinetics of cell death in neuronal cell lines, whereas this method is less suitable for the assessment of neuronal differentiation and viability of primary neurons.
神经元细胞死亡的检测是神经退行性疾病细胞培养模型的标准要求。虽然有大量的生存能力检测方法可用于体外应用，但这些方法大多数都是终点测量方法，只能提供关于细胞死亡动力学的少量信息。在这里，我们验证了基于阻抗测量的 xCELLigence 系统，用于实时检测永生海马神经元（HT-22 细胞）、神经元祖细胞（NPC）和分化的原代皮质神经元中的细胞死亡。我们发现，在 HT-22 细胞和 NPC 中，阻抗测量与终点生存能力检测之间存在良好的相关性，可用于检测增殖、细胞死亡动力学以及凋亡的药理学抑制剂的神经保护作用。在原代神经元中，我们无法检测到细胞分化过程中的树突生长。xCELLigence 系统可以检测原代神经元中的细胞死亡，然而，基于阻抗测量的细胞指数变化在很大程度上取决于损伤的严重程度。离子霉素诱导的细胞死亡，例如，表现为一个快速的过程，涉及到强烈的细胞解体，这使得基于阻抗的检测成为可能。然而，由谷氨酸诱导的细胞死亡等伴有不太明显的形态变化的细胞死亡，这种方法则不容易检测到。总之，我们的数据表明，阻抗测量是检测神经元细胞系中增殖和细胞死亡动力学的一种方便可靠的方法，而这种方法不太适合评估神经元分化和原代神经元的活力。
