To evaluate the adhesion of ADSCs on the DC-ECM and S/D scaffolds, scaffolds were first transferred to 12-well plates, then 50 μL of a suspension containing 5×105 ADSCs was seeded into each scaffold, and plates were incubated in a 5% CO2 humidified incubator at 37°C for 2 h, then 3 mL of medium was added to each well and plates were transferred back to the incubator for 7 days of culture. Medium was changed every 3 days. After 7 days of culturing, scaffolds with cells were fixed with 2.5% glutaraldehyde at 4°C for 12 h, then dehydrated through a graded ethanol series. The dried scaffolds were cut into cross sections of 1-mm thick and coated with gold. The adhesion of ADSCs on the scaffolds was observed by SEM.
The viability of the ADSCs within the scaffolds was assessed by use of a live/dead cell viability assay kit (Abcam, Cambridge, UK) as per the manufacturer’s instruction after 7 days of culture. After incubation with the live/dead staining solution for 30 min, live cells (green) and dead cells (red) were observed by confocal microscopy (Leica < Heidelberg, Germany).
The viability of the ADSCs within the scaffolds was assessed by use of a live/dead cell viability assay kit (Abcam, Cambridge, UK) as per the manufacturer’s instruction after 7 days of culture. After incubation with the live/dead staining solution for 30 min, live cells (green) and dead cells (red) were observed by confocal microscopy (Leica < Heidelberg, Germany).