Characterization of BMSC surface markers

To identify the cell surface markers of BMSCs, P4 cells were collected according to the previous literature, detached from plates with 0.25% trypsin-EDTA solution, and gently resuspended in flow cytometry staining solution at a density of 5 × 10⁶ cells/mL [3 (link), 24 (link)]. Then, the following antibodies were applied for 30 min at room temperature protected from light: anti-CD29-PE, anti-CD34-PE, anti-CD44-PE, anti-CD45-PE, anti-CD73-PE, and anti-CD105-PE. The nonspecific Fc receptor was blocked with anti-CD16/32 antibodies. Nonspecific fluorescence was determined by incubation with isotype-matched anti-mouse monoclonal antibodies. After staining, the samples were processed with EPICS XL-MCL (Beckman Coulter, Brea, CA, USA), and the analysis was performed with EXPO32 ADC.