Article
Introduction of a cell culture device for electrical stimulation of spiral ganglion cells
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Published: | September 22, 2005 |
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Deafness leads to hair cell loss followed by the loss of spiral ganglion cells. Electrical stimulation of spiral ganglion cells has shown to be effective for spiral ganglion cell protection following deafness (Kanzaki et al., 2002). However, it is still unknown what exactly the most effective stimulation conditions are to protect spiral ganglion cells. Therefore we wanted to build a device to rapidly test cultured spiral ganglion cells in the presence of electrical fields (AC).
The device was first tested for pH and temperature behaviour of the culture media under stimulation conditions. Initial experiments with fibroblasts (NHDF) were carried out using stimulus trains of 50 biphasic rectangular pulses (10ms, 50Hz) at constant voltage (10-60V) followed by an interburst interval of 19 s. After cultivation fibroblasts were harvested, stained with propidium iodide and analyzed by flow cytometry (FACS). Additionally the electrical field strength in the cultivation chambers was determined under the different stimulation conditions.
During electrical stimulation the pH of the culture media remained constant at pH 7,4 whereas the temperature of the culture media remained in acceptable conditions for cell cultivation purposes (37°C) at voltages of up to 40 V (480V/m in cultivation chamber). Cultivated and electrically stimulated fibroblasts showed no increased rate of damage when compared to unstimulated fibroblasts at voltages of up to 40V.
These results demonstrate that the developed cell culture device is suitable for experiments analysing the effects of an electrical stimulation on cultivated cells, especially spiral ganglion cells.