Replication Data for: beta-galactosidase of "Replicative Senescence in Mesenchymal Stem Cells: An In Vitro Study on Mitochondrial Dynamics and Metabolic Alterations"

To evaluate Senescence-Associated β-galactosidase (SA-β-gal) bMSCs were seeded on a 24-well plate (Costar, Sigma-Aldrich, St. Louis, MO, USA). Cells were then washed with phosphate-buffered saline (PBS), fixed in 2% formaldehyde and 0.2% glutaraldehyde for 10–15 min at room temperature, and subsequently incubated overnight at 37 °C with a freshly prepared staining solution containing 1 mg/mL X-Gal (B4252, Sigma-Aldrich), 40 mM citric acid/sodium phosphate buffer (pH 6.0), 5 mM potassium ferrocyanide, 5 mM potassium ferricyanide, 150 mM NaCl, and 2 mM MgCl2. The following day, cells were rinsed with PBS and the blue-colored precipitates were quantified at 650 nm using the Varioskan LUX Multimode Microplate Reader (Thermo Fisher Scientific, Waltham, MA, USA).