Anti cd29 apc

BMMSCs were isolated from the femurs and tibias of sham-operated mice or OVX mice treated with PBS, TDN_MCP-1, TDN_FasL, or TDNs. Femurs and tibias were dissected out and cleaned of connective tissue. Cells were flushed out from long bones by a syringe with α-MEM supplemented with 20% FBS, 2 mM l-glutamine (Sigma-Aldrich, USA), and 1% penicillin/streptomycin (Invitrogen, USA). Single-cell suspensions were seeded in dishes and initially maintained in an atmosphere of 5% CO₂ at 37 °C. The medium was changed every 3 days. After reaching 80% confluence, cells were passaged using 0.25% trypsin.
Cell surface markers of BMMSCs were detected by flow cytometry analysis. Cells were harvested and washed in PBS. Then, the single-cell suspension was incubated with mouse anti-CD105-APC (BioLegend, 120413), anti-CD11b-FITC (BioLegend, 101206), anti-CD45-PE (eBioscience, 12-0451), anti-CD29-APC (eBioscience, 17-0291-82) and anti-CD34-PE (BioLegend, 119307) antibodies. Finally, cells were washed twice in PBS, subjected to flow cytometry analysis with a flow cytometer (Cytomics FC 500; Beckman-Coulter) and analyzed with FlowJo_V10 software.