Impedance-based monitoring of ongoing cardiomyocyte death induced by tumor necrosis factor-alpha.
Deregulated cardiomyocyte death is a critical risk factor in a variety of cardiovascular diseases. Although various assays have been developed to detect cell responses during cell death, the capability of monitoring cell detachment will enhance the understanding of death processes by providing instant information at its early phase. In this work, we developed an impedance-sensing assay for real-time monitoring of cardiomyocyte death induced by tumor necrosis factor-alpha based on recording the change in cardiomyocyte adhesion to extracellular matrix. Electrochemical impedance spectroscopy was employed in impedance data processing, followed by calibration with the electrical cell-substrate impedance-sensing technique. The adhesion profile of cardiomyocytes undergoing cell death processes was recorded as the time course of equivalent cell-substrate distance. The cell detachment was detected with our assay and proved related to cell death in the following experiments, indicating its advantage against the conventional assays, such as Trypan blue exclusion. An optimal concentration of tumor necrosis factor-alpha (20 ng/mL) was determined to induce cardiomyocyte apoptosis rather than the combinative cell death of necrosis and apoptosis by comparing the concentration-related adhesion profiles. The cardiomyocytes undergoing apoptosis experienced an increase of cell-substrate distance from 59.1 to 89.2 nm within 24 h. The early change of cell adhesion was proved related to cardiomyocyte apoptosis in the following TUNEL test at t = 24 h, which suggested the possibility of early and noninvasive detection of cardiomyocyte apoptosis.
Qiu
Yiling
Y
Laboratory for Microsystems Technology, Department of Mechanical Engineering, Boston University, Boston, Massachusetts, USA.
Liao
Ronglih
R
Zhang
Xin
X
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D016923:Cell Death / Q000502:physiology*; D002470:Cell Survival; D017097:Electric Impedance; D005109:Extracellular Matrix / Q000502:physiology; D020287:In Situ Nick-End Labeling; D008297:Male; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D013057:Spectrum Analysis / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D014409:Tumor Necrosis Factor-alpha