Characterization of Three-Dimensional Tissue Cultures Using Electrical Impedance Spectroscopy

AbstractElectrical impedance spectroscopy was used to characterize the cell environment of multilayered cell cultures (MCCs), a culture system in which cells are grown on a permeable support membrane to form a thick disc of cells with tumor-like properties. Cultures were grown using SiHa tumor cells as well as V79 wild-type cells and V79/DOX cells cultivated to exhibit multidrug resistance. Electrical impedance measurements were made on MCCs over a frequency range of 0.1kHz to 1MHz. Data analysis using a simple electrical model for the cell environment yielded estimates for parameters related to the intra- and extracellular resistance and net membrane capacitance, which were then related to MCC thickness. The extracellular fraction and tortuosity of the MCCs were determined in separate experiments where the rate of diffusion and the equilibrium level of C14-inulin, which does not penetrate the cell membrane, was measured within MCCs. Impedance measurements predicted the barrier to diffusion posed by the extracellular space of MCCs to be roughly two times greater than that inferred from the C14-inulin experiments. However, the relative ranking of the three cell types used to grow MCCs was similar for the two methods. Results indicate that impedance spectroscopy is well suited for use in characterizing the MCC cell environment, offering a fast, nondestructive method for monitoring cell culture growth and integrity