CHO cell dysfunction due to radiation-induced bystander signals observed by real-time electrical impedance measurement.

Radiation-induced bystander effects (RIBE) have raised many concerns about radiation safety and protection. In RIBE, unirradiated cells receive signals from irradiated cells and exhibit irradiation effects. Until now, most RIBE studies have been based on morphological and biochemical characterization. However, research on the impact of RIBE on biophysical properties of cells has been lagging. Non-invasive indium tin oxide (ITO)-based impedance systems have been used as bioimpedance sensors for monitoring cell behaviors. This powerful technique has not been applied to RIBE research. In this work, we employed an electrical cell-ITO substrate impedance system (ECIIS) to study the RIBE on Chinese hamster ovary (CHO) cells. The bioimpedance of bystander CHO cells (BCHO), alpha(α)-particle (Am-241) irradiated CHO (ICHO), and untreated/unirradiated CHO (UCHO) cells were monitored with a sampling interval of 8 s over a period of 24 h. Media from ICHO cells exposed to different radiation doses (0.3 nGy, 0.5 nGy, and 0.7 nGy) were used to investigate the radiation dose dependence of BCHO cells' impedance. In parallel, we imaged the cells at times where impedance changes were observed. By analyzing the changes in absolute impedance and cell size/cell number with time, we observed that BCHO cells mimicked ICHO cells in terms of modification in cell morphology and proliferation rate. Furthermore, these effects appeared to be time-dependent and inversely proportional to the radiation dose. Hence, this approach allows a label-free study of cellular responses to RIBE with high sensitivity and temporal resolution and can provide crucial insights into the RIBE mechanism.