Quantifying MSC Migration Dynamics

The procedure of cell migration assay was followed in a previous study [25 (link)]. First, 1 × 10⁴ per well of MSCs were added into Oris seeding stoppers for 24 h of incubation to reach confluency. Next, the stoppers were removed and used as a pre-migration reference (t = 0 h). The post migration was represented at 24 h. Next, 200 μL of a Calcein AM solution (2 μM, Sigma-Aldrich, Burlington, MA, USA) was added to the culture plates and stained for 30 min at the time points of 0 and 24 h. The cells were observed by a Zeiss Axio Imager A1 fluorescence microscope (White Plains, NY, USA). Cell migration distance was semi-quantified using Image J 5.0 software (Becton Dickinson, Canton, MA, USA). The MSC alone represented as the control.