Anti rat cd29 fitc

Bone marrow-derived MSCs were isolated from 2-week-old Sprague-Dawley (SD) rats as previously described [10 (link)]. In brief, femurs and tibias from 2-week-old rats were used to collect the total bone marrow and were then subjected to treatment with red blood cell lysis buffer two times for 5 min at room temperature and washed with PBS. Afterwards, the cells were cultured in Dulbecco’s modified Eagle medium: nutrient mixture F-12 (DMEM/F12) with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37 °C with 5% CO₂. After 4–6 days of culturing, spindle cells reached 80% confluence and were passaged. Passage 3 cells were identified by flow cytometry with the following antibodies: anti-rat CD105-PE (12-1051, eBioscience), anti-rat CD73-PE (551123, BD), anti-rat CD90-PE (551401, BD), anti-rat CD29-FITC (555005, BD), anti-rat CD45-PE (6828, BD), and anti-rat CD11b/c-FITC (b141556, Biolegend). Passage 3 cells were used for the following experiments.

Rat MSCs (rMSCs) were harvested from the bone marrow of healthy SD rats (80‐100 g) and cultured as described previously.¹² Briefly, the rats were killed by an intraperitoneal injection of pentobarbital sodium (30 mg/kg bodyweight), and bone marrow cells were aspirated from the femurs and tibias. Then, cells were plated in 25‐cm² flasks with Dulbecco's modified Eagle's medium (HyClone) containing 15% foetal bovine serum and 1% penicillin‐streptomycin, at 37°C. After 72 h, the medium was replaced with fresh medium, and the adherent cells were defined as passage 0 MSCs. The surface markers of MSCs were identified by flow cytometric analysis using fluorophore‐conjugated antibodies: anti‐rat CD29‐FITC, CD44‐FITC, CD34‐PE and CD45‐PE (BD Biosciences, Franklin Lakes, NJ, USA). Cells at passages 3 or 4 were used for experiments.