Real-Time Monitoring of hBM-MSC Proliferation

The effects of the UBC gene KD and transfection on hBM-MSC proliferation were monitored using xCELLigence RTCA DP system (Roche), which allows for label-free, real-time monitoring of the cultured cell viability using impedance^{34 (link)}. A total of 50 μL of antibiotic-free basic cell culture medium was added to an E-plate with plate bottom coated with gold electrodes. After incubation at room temperature for 15 min, the background intensity was measured. Cells (n = 10,000) were suspended in 150 μL of antibiotic-free basic cell culture medium and seeded in each well of the E-plate. Twenty-four hours after E-plate was installed in the xCELLigence system, UBC gene KD was performed combined with control treatments. The cell index was measured every 10 min, which is a relative values calculated by analyzer. Data analysis was carried out using RTCA software 2.0 supplied with the instrument. All experiments were performed in triplicate.