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Three dimensional (3D) stem cell culture has recently received considerable attention because it may enable the development of in vitro 3D tissue models. In particular, label-free and real-time monitoring of stem cell differentiation is of importance for tissue engineering applications; however, only a few non-invasive monitoring methods are available, especially for 3D cell culture. Here, we describe impedance cell sensors that allowed the monitoring of cellular behaviors in 2D and 3D cell cultures in real-time. Specifically, apparent capacitance peaks appeared in both 2D and 3D cell culture systems when human mesenchymal stem cells (hMSCs) were cultured in osteogenic induction medium. In contrast, when hMSCs were cultured in adipogenic induction medium, the capacitance increased monotonically. In addition, distinct characteristics were noted in the plots of capacitance versus conductance for the cells cultured in osteogenic and adipocyte induction media. These results demonstrated that the differentiation of hMSCs toward osteoblasts and adipocytes in 2D and 3D cell culture systems could be discriminated non-invasively by measuring the real-time capacitance and conductance. Furthermore, the vertical distribution of cellular activities in 3D cell cultures could be monitored in real-time using the 3D impedance cell sensors. Thus, these sensors may be suitable for monitoring the differentiation of various stem cells into different types of cells with distinct dielectric properties for tissue engineering applications.

Label-free and real-time monitoring of human mesenchymal stem cell differentiation in 2D and 3D cell culture systems using impedance cell sensors