MSCs were isolated from the bone marrow of 4-week-old rats. The experimental rats were euthanized via an intraperitoneal injection of sodium pentobarbital, and the femur and tibia were separated under sterile conditions.10 (link) The marrow cavity was washed repeatedly with Dulbecco’s Modified Eagle’s Medium (DMEM; GIBCO, Grand Island, NY, USA). The cell suspension was centrifuged at 350 g for 10 min, and the supernatant was discarded. The precipitate was mixed with erythrolysis buffer (Solarbio, Beijing, China) for 3 min. Cells were subjected to centrifugation and washing, followed by culture in DMEM supplemented with 10% fetal bovine serum.
The specific antibody markers of MSCs were detected using flow cytometry. Positive antibody markers included CD90-FITC (eBioscience, San Diego, CA, USA) and CD105-FITC (Abcam, Cambridge, UK). Negative markers were CD34 (Abcam) combined with goat anti-rabbit IgG (Abcam) labeled with phycoerythrin and FITC-CD45. Cells stained with an isotype control (IgGFITC) were used as the negative control.
The specific antibody markers of MSCs were detected using flow cytometry. Positive antibody markers included CD90-FITC (eBioscience, San Diego, CA, USA) and CD105-FITC (Abcam, Cambridge, UK). Negative markers were CD34 (Abcam) combined with goat anti-rabbit IgG (Abcam) labeled with phycoerythrin and FITC-CD45. Cells stained with an isotype control (IgGFITC) were used as the negative control.
