Real-time Cellular Impedance Monitoring and Imaging of Biological Barriers in a dual flow membrane bioreactor

Abstract: The generation of physiologically relevant in-vitro models of biological barriers can play a key role in understanding human diseases and in the development of more predictive methods for assessing toxicity and drug or nutrient absorption. Here, we present an advanced cell culture system able to mimic the dynamic environment of biological barriers while monitoring cell behaviour through real-time impedance measurements and imaging. It consists of a fluidic device with an apical and a basal flow compartment separated by a semi-permeable membrane. The main features of the device are the integration of a transepithelial electrical impedance (TEEI) meter and transparent windows for optical monitoring within a dual flow system. Caco-2 cells were cultured in the TEEI bioreactor under both flow and static conditions. Although no differences in the expression of peripheral actin and occludin were visible, the cells in dynamic conditions developed higher impedance values at low frequencies, indicative of a higher paracellular electrical impedance and thus suggesting accelerated barrier and tight junction formation with respect to the static cultures. TEEI measurements at high frequency also enabled monitoring the evolution of transcellular impedance during culture. The cells subject to flow showed a typical RC behaviour, while the controls showed minimal capacitive behaviour, again highlighting the differences between flow and static conditions.