Phenotypic identification of BMSCs

Before in vivo experiments, the mesenchymal phenotype of the cells was assessed by fluorescence-activated cell sorting (FACS). The cells in the third passage were harvested with 0.25% trypsin/EDTA, resuspended at a concentration of 1 × 10⁶ cells/mL in PBS, and separated in a microcentrifuge tube. According to the instructions of the manufacturer, corresponding doses of CD45, CD29, and CD90 monoclonal antibodies were added to each tube in turn. Meanwhile, the same type of negative control was established. Then, it was mixed well and incubated at 4 °C for 1 h and detected by flow cytometry.