Abstract
Purpose: Poor survival rate of mesenchymal stem cells (MSCs) following their transplantation is one of the major challenges in their therapeutic application. Therefore, it is necessary to augment the viability of the MSCs in order to improve their therapeutic efficacy. Several strategies have been used to overcome this problem. Preconditioning of MSCs with oxidative stresses has gained a lot of attention. Therefore, in the present study, we investigated the effects of simultaneous preconditioning of MSCs with hydrogen peroxide and serum deprivation stresses on their survival and resistance to stressful conditions.Methods: MSCs were isolated from human umbilical cord blood. To perform simultaneous preconditioning, the cells were cultured in DMEM medium containing 1, 2.5 and 5 percent FBS and different concentrations of H2O2 (5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 80 and 100 µM) for 24 hrs. Then, the cells were cultured in recovery culture medium. Finally, one group of the cells was exposed to a lethal concentration of H2O2 (300µM), and the other cells were cultivated in FBS free DMEM medium as the lethal situation. In addition, the percentage of apoptotic cells was analyzed using Caspase 3 assay kit.Results: Simultaneous preconditioning of the MSCs with 15µM H2O2 plus serum deprivation, 2.5% FBS, significantly increased the resistance of the cells to the toxicity induced following their cultivation in FBS free DMEM medium. It exerted the protective effect on the cells after treating with the lethal dose of H2O2 as well.Conclusion: Simultaneous preconditioning of MSCs with oxidative and serum deprivation stresses enhances their survival against harsh conditions, which might increase the viability and stability of the MSCs following their transplantation.
Highlights
One group of the cells was exposed to a lethal concentration of H2O2 (300μM), and the other cells were cultivated in fetal bovine serum (FBS) free Dulbecco’s modified eagle medium (DMEM) medium as the lethal situation
Mononuclear cells were separated by Ficoll-Hypaque density gradient centrifugation, and seeded cell culture flasks containing low glucose Dulbecco’s modified eagle medium (DMEM), antibiotics (0.01% penicillin/ streptomycin) and 10% fetal bovine serum (FBS) (All of materials purchased from Gibco, Germany)
H2O2-preconditioning enhanced cell survival of UCBMSCs and decreased their apoptosis rate As described in above, UCB-mesenchymal stem cells (MSCs) were treated by different concentration of H2O2
Summary
It has been clear that the mesenchymal stem cells (MSCs) are promising cell source for the treatment of a variety of human diseases[1,2] including severe aplastic anemia[3,4] acute graft- versus-host disease,[5] cardiovascular diseases, acute liver failure[6] and kidney injuries.[7,8] Multilineage differentiation potential, immune modulatory properties and ability to localize to injured sites have made MSCs as an appropriate alternative.[2,8] According to animal and clinical studies, MSC transplantation can restore cardiac function probably by myogenesis and angiogenesis after myocardial infarction.[9,10,11] there has been an increasing interest in using these cells in cell therapy but one of the main obstacles of their application is poor survival after transplantation. It has been shown that a majority of implanted cells die within few days after transplantation.[12,13] Endogenous and environmental factors[14,15,16,17,18] including inflammatory responses, lack of nutritional factors, hypoxia, reactive oxygen species (ROS) such as superoxide anion (O2-), hydroxyl radical (OH-), and hydrogen peroxide (H2O2), induce apoptosis and higher cell death either in vitro or in vivo MSCs microenvironments specially in ischemic heart medium.[16]
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