Characterization of Rat Bone Marrow Mesenchymal Stem Cells

Rat BM-MSCs were detached, collected, and centrifuged at 300 × g for 5 min and then suspended in the expansion medium. The viability of BM-MSCS was evaluated in an aliquot using Trypan blue stain by counting the stained live cells and excluding the unstained dead cells as reported by Al-Mutairi et al. [25 ]. For BM-MSC, surface antigen phenotypic analysis and cell markers were assessed [26 (link)]. After being washed twice with phosphate-buffered saline (PBS, PH 7.4; 137-mM NaCl, 2.7-mM KCl, 10-mM Na₂HPO₄, and 1.8-mM KH₂PO₄) (Lonza, Germany) containing 1% bovine serum albumin (ALB) (Sigma-Aldrich), 0.2×10⁶ cells were stained with anti-CD34, anti-CD45, anti-CD29, anti-CD73, and anti-CD90 antibodies (BD Biosciences, USA). A FACSCalibur flow cytometer (BD Biosciences) was used to examine conjugated cells as well as determine the proportions of positive and negative populations. A negative sample with untreated isotype cells was used as a control.