Single-cell impedance analysis of osteogenic differentiation by droplet-based microfluidics

Single-cell analysis is critical to understanding its heterogeneity and biological processes, such as stem cell differentiation, and elucidating the underlying mechanisms of cellular metabolism. New tools to promote intercellular variability studies help elucidate cellular regulation mechanisms. Here an impedance measurement and analysis system was built to monitor the osteogenic differentiation of single bone marrow mesenchymal stem cells (BM-MSCs) in droplets. The biochip including a microelectrode array was designed based on droplet microfluidics and fabricated. A novel theoretical electrical model was proposed to simulate the electrical properties of cells in the droplets. Impedance measurements showed that single cells are substantially heterogeneous during osteoblast differentiation at different stages (days 0, 7, 14 and 21) and different cell passages (passages 6, 7 and 11). This result was consistent with the appearance of two biomarkers (alkaline phosphatase and calcium nodules), which are the gold standard biomarkers of osteoblastogenesis and differentiation. The device enabled highly efficient single-cell trapping, accurate positioning, and sensitive, label-free and noninvasive impedance measurements of individual cells with multiple channels. This system provides a strategy for exploring the processes of osteoblastogenesis and differentiation at the single-cell level and has substantial potential for applications in the biomedical field.