BM-MSCs were isolated as described previously [11]. Briefly, the bone marrow of euthanized 3-week-old male SD rats was obtained by flushing the marrow cavity of their femurs with low-glucose DMEM (Gibco) supplemented with 10% FBS (Gibco) and 1% streptomycin-penicillin solution (Gibco). After centrifugation, the cells were resuspended and cultured at 5% СО2 and 37°C in a humidified atmosphere. After 2 days, the nonadherent cells were removed, and the medium was changed every 2–3 days. BM-MSCs at the third passage were characterized using flow cytometry analysis, and BM-MSCs from the third and fourth passages were used for the following experiments. The presence of the surface markers of BM-MSCs (CD29, CD34, CD45, and CD90) was verified using flow cytometry with the appropriate primary labeled antibodies (Abcam, Cambridge, UK). The differentiation of osteogenic cells and fat cells for BM-MSCs was evaluated by Alizarin red staining and oil red staining, respectively.

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