Cd90 fitc ab226

Rat bone marrow-derived MSCs were obtained from the femur and tibia of healthy male Sprague Dawley rats (100-120 g). These cells were cultured in low glucose DMEM containing 10% FBS (Gibco, Invitrogen, New York, USA), 2.5 mM L-glutamine, and penicillin/streptomycin (Gibco, Invitrogen, New York, USA) in a 5% CO 2 incubator at 37 °C. When the cultures reached 90% con uence, MSCs were harvested using 0.25% trypsin-EDTA (Thermo Fisher Scienti c, Waltham, MA, USA). MSCs were subcultured at the ratio of 1:3.
For all experiments, MSCs (passages 3-4) were collected and washed with sterile saline to achieve a single cell suspension for model treatment or analysis. To identify MSCs, the cells were incubated with the following antibodies against surface antigens (Abcam,UK): CD90-FITC (ab226), CD29-PE/Cy7 (ab95622), CD45-FITC (ab33916) and CD34-FITC (11-0341-82, eBioscience, USA). Finally, MSCs were washed and resuspended in 0.4 mL of PBS for analysis using a BD FACSCalibur ow cytometer and Cell Quest software (BD Biosciences, San Jose, CA, USA). Rat MSCs induction and differentiation were carried out using Oil red O staining to verify adipogenesis, using Alizarin red staining to con rm osteogenesis.