Impedance measurement for real time detection of neuronal cell death

• Published in: Journal of neuroscience methods 2012-01, Vol.203 (1), p.69-77
• Main Authors: Diemert, S., Dolga, A.M., Tobaben, S., Grohm, J., Pfeifer, S., Oexler, E., Culmsee, C.
• Format: Article
• Language: English
• Subjects: Apoptosis; Cell Death - drug effects; Cell Death - physiology; Cell Line; Electric Impedance; Glutamic Acid - toxicity; Humans; Impedance measurement; Neuronal cell culture; Neuronal cell death; Neurons - drug effects; Neurons - pathology; Neuroprotective Agents - pharmacology; Real time viability assay; xCELLigence
• ISSN: 0165-0270; 1872-678X
• DOI: 10.1016/j.jneumeth.2011.09.012

► Real time monitoring of neuronal cell death in cell lines and primary neurons by impedance measurements. ► For neuronal cell lines and neuronal progenitor cells impedance readouts correlate well with viability assays. ► The kinetics of cell death in cell lines is well reflected by impedance measurements. ► Monitoring cell death in primary cultures is less accurate compared to cell lines. 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.