β galactosidase cell staining kit

Senescence was analyzed using a β-Galactosidase Cell Staining Kit (Cell Signaling Technology, Inc., USA) according to the manufacturer's instructions. In addition to negative control cells, senescence was induced in untreated AD-hMSCs by cultivation in high-glucose medium (4.5 g/L) for 21 days.^{35 (link)}

Senescence was evaluated using a β-galactosidase cell staining kit (Cell Signaling, Danvers, MA, USA) as recommended by the manufacturer. Briefly, BEAS-2B cells were cultured in 3.5 cm dishes were rinsed, fixed with 1 × fixative solution for 15 min, and washed twice with PBS. β-galactosidase staining solution (930 μL 1 × staining solution with 10 μL supplement A, 10 μL supplement B, and 50 μL 20 mg/mL X-gal in DMF) was added, and the cells were incubated at 37 °C without CO₂ overnight. Images were taken with a Nikon LED microscope the following day, as previously described [35 (link)]. Experiments were repeated two times.