Combined preconditioning with hypoxia and Hydrogen Peroxide Improved efficiency of Mesenchymal Stem Cell in cell culture

Preconditioning with hypoxia or ROS such as hydrogen peroxidase improved long term survival of Mesenchymal Stem Cells. But combined therapy with both of them is not investigated enough in cell culture. Therefore, the present study investigates the protective effects of co-preconditioning of the cells with H2O2 and hypoxia in cell culture. Bone Marrow MSCs were cultured and divided in 6 groups; control, groups I and II exposed to H2O2, group III treated with COCL2, Groups IV and V treated withbothH2O2andCOCL2 during 6, 12, 24 and 48h. Then, all groups were underwent to lethal dose of H2O2 (300 μM) for 24 hours followed by 24h recovery. Afterward, MTT assay and trypan blue staining were conducted to evaluate the cell proliferation and viability. Also, TUNEL was done to study the cell apoptosis. Cell proliferation and viability after preconditioning with H2O2 and hypoxia were considerably increased compared to the control group after 6, 12, 24 and 48 hours (P < 0.01). Furthermore, cell proliferation and viability in groups IV and V were remarkably more than control and others preconditioning groups (P < 0.01). Apoptosis was significantly decreased in preconditioned groups compared to the control group after 12 (P < 0.01), 24 (P < 0.05) and 48 hours (P < 0.01). Also, cell apoptosis was decreased significantly in groups IV and V after 12, 24 and 48 hours compared to control and other preconditioning groups (P < 0.01). This study clearly demonstrated that preconditioning with H2O2 and COCL2 can improve BMMSCs proliferation and viability and also have inhibitory effect on cell apoptosis.

Keywords: Bone Marrow Mesenchymal Stem Cells, hypoxia, oxidative stress, co-preconditioning.

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