Preconditioning and post-preconditioning states of mesenchymal stem cells with deferoxamine: A comprehensive analysis

Abstract

Mesenchymal stem cells (MSCs) derive their therapeutic potential from their secretomes, which can be modulated by external stimuli. Hypoxia is one such stimulus, and research on preconditioning MSCs with hypoxia-mimetic agents is rising. However, the effects of these preconditioning processes and the resulting metabolic status require further investigation. This study evaluated the effects of deferoxamine (DFX), a hypoxia-mimetic agent, preconditioning on MSCs in serum and serum-free environments. The influence of hypoxia on cell metabolism was examined during and after preconditioning by assessing cytotoxicity, proliferation, migration, secretomes, senescence, autophagy, and apoptosis mechanisms. The optimal DFX dose and duration for preconditioning were determined as 150 μM and 24 h based on cytotoxicity testing. Accordingly, DFX preconditioning significantly upregulated HIF-1α expression, increasing protein secretion and reducing total oxidant status. DFX appears to enhance the therapeutic potential of MSCs by increasing their secretome and antioxidant capacity. However, upon DFX removal, HIF-1α levels returned to normal, and the associated positive effects diminished. Autophagy was markedly enhanced during DFX preconditioning, potentially improving metabolic activity, preserving cellular integrity, and preparing MSCs for ischemic environments. Autophagy returned to baseline after DFX withdrawal, indicating a temporary hypoxia-mimetic response. In a serum-containing medium, specific effects of preconditioning were relatively weak to be observed. This study demonstrates that DFX-preconditioning increases MSCs' metabolic activity and enhances their adaptive cellular response. However, the effect may be transient, which provides insights into the behavior of MSCs in ischemic environments and emphasizes the need to evaluate the long-term effects of hypoxia-mimetic agents.