H₂O₂ Damages the Stemness of Rat Bone Marrow-Derived Mesenchymal Stem Cells: Developing a "Stemness Loss" Model

BACKGROUND The number of patients with spinal cord injury caused by motor vehicle accidents, violent injuries, and other types of trauma increases year by year, and bone marrow mesenchymal stem cell (BMSC) transplants are being widely investigated to treat this condition. However, the success rate of BMSCs transplants is relatively low due to the presence of oxidative stress in the new microenvironment. Our main goals in the present study were to evaluate the damaging effects of H₂O₂ on BMSCs and to develop a model of "stemness loss" using rat BMSCs. MATERIAL AND METHODS Bone marrow-derived mesenchymal stem cells were obtained from the bone marrow of young rats reared under sterile conditions. The stem cells were used after 2 passages following phenotypic identification. BMSCs were divided into 4 groups to evaluate the damaging effects of H₂O₂: A. blank control; B. 100 uM H₂O₂; C. 200 uM H₂O₂ and D. 300 uM H₂O₂. The ability of the BMSCs to differentiate into 3 cell lineages and their colony formation and migration capacities were analyzed by gene expression, colony formation, and scratch assays. RESULTS The cells we obtained complied with international stem cell standards demonstrated by their ability to differentiate into 3 cell lineages. We found that 200-300 uM H₂O₂ had a significant effect on the biological behavior of BMSCs, including their ability to differentiate into 3 cell lineages, the expression of stemness-related proteins, and their migration and colony formation capacities. CONCLUSIONS H₂O₂ can damage the stemness ability of BMSCs at a concentration of 200-300 uM.