Pe conjugated cd44

The surface markers of isolated cells were analyzed by flow cytometry analysis. Briefly, the isolated cells (1 × 10⁶ cells, passage 3) from the three dogs were, respectively, suspended in 100 μL phosphate-buffered saline (PBS) containing 10 μg/mL antibodies for PE-conjugated CD29 (303003, BioLegend, USA), PE-conjugated CD44 (103024, BioLegend, USA), PE-conjugated CD90 (561970, BD Biosciences, USA), PE-conjugated CD105 (bs-0579R-PE, Bioss, CHN), FITC-conjugated CD73 (bs-23233R-FITC, Bioss, CHN), PE-conjugated CD34 (559369, BD Biosciences, USA), and FITC-conjugated CD45 (11-5450-42, eBioscience, USA). After incubation for 30 min at 4°C, the cells were washed with PBS and then resuspended in 500 μL of PBS for analysis. As for CD11b, the isolated cells (1 × 10⁶ cells, passage 3) were suspended in 100 μL phosphate-buffered saline (PBS) containing 10 μg/mL CD11b (MA5-16604, eBioscience, USA). After incubation for 30 min at 4°C, the cells were washed with PBS and then labeled with goat antirabbit IgG (H + L) cross-adsorbed secondary antibody, FITC (F-2765, Invitrogen), at a dilution of 1 : 500 for 1 h at room temperature. Cell fluorescence was evaluated by flow cytometry using a DxP Athena™ flow cytometry system (Cytek) and analyzed with FlowJo 10 software (Tree Star, USA).