Real-Time Monitoring of the Formation and Culture of Hybrid Cell-Microbiomaterial Spheroids Using Non-Faradaic Electrical Impedance Spectroscopy.

Cellular spheroids are considered a popular option for modeling healthy and diseased tissues in vitro and as injectable therapies. The formation and culture of spheroids can make use of different three-dimensional (3D) culture platforms, but the spheroids' analysis often has to rely on endpoint assays. In this study, we propose a microfluidic bioreactor to culture and nondestructively monitor human mesenchymal stem cell (hMSC) spheroids over time using non-Faradaic electr(ochem)ical impedance spectroscopy (EIS). For this, an array of porous microwells thermoformed from ion track-etched thin films and a pair of sensing electrodes from transparent indium tin oxide are integrated into the flow and culture chamber of the bioreactor. To measure the spheroid's electrical properties, the electrodes are connected to a frequency response analyzer (FRA), with a multiplexer in between to enable the operation of more than one bioreactor at the FRA at the same time. We find differences between the complex resistance/impedance and/or capacitance data of a reference condition without cells, a two-dimensional (2D) hMSC culture, hMSC spheroids, and hybrid spheroids aggregated from hMSCs and titanium or hydroxyapatite microparticles. We also found differences between different culture durations. These results suggest that our device can sense the presence and spatial arrangement of cells and micro(sized) biomaterials as a function of time.