Real-Time Cell Adhesion and Proliferation

The xCELLigence system (RTCA-DP version; Roche Diagnostics, Mannheim, Germany) was used to determine the adhesion and proliferation of PE-DMBSCs and DBMSCs after preconditioning, and in DBMSCs after HMOX1 inhibition. Briefly, 100 μL complete medium was added to each well in 16-well culture plate, E-Plate 16 (catalog number 05469813001, Roche Diagnostics), and the background impedance was achieved as previously described [29 (link)]. Treated and untreated control cells were seeded in quadruplicate wells, and equilibrium was achieved by leaving the culture plates for 30 min at RT before data recording. The culture plates were placed in the xCELLigence system at 37 °C in a cell culture incubator and cell index was monitored for 72 h. Data was analyzed by the RTCA xCELLigence software (version 1.2.1). For cell adhesion, data was measured after 2 h, whereas the rate of cell proliferation was calculated after 24, 48, and 72 h and after normalization with the adhesion data. Data is expressed as cell index with mean and standard errors. All experiments were performed in triplicate.