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Role of intracellular level of glutathione on the therapeutic capacity of human mesenchymal stem cell Jisun Lim, YongHwan Kim, Seungun Lee, Jinbeom Heo, Hye-Yeon Lee, Hyein Ju & DongMyung Shin Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea Continuous exposure to oxidative stress during the expansion of mesenchymal stem cells (MSCs) based on traditional culture techniques results in a progressive loss of proliferative and differentiation potential. Glutathione (GSH) is an important antioxidant in mammalian cells, preventing damage to important cellular components caused by reactive oxygen species (ROS). Here, we show that the MSCs with high content of intracellular GSH have the enhanced therapeutic potency of MSCs. Employing a newly synthesized fluorescent probe (FreSH tracer) which exhibit reversible reaction with various thiol-compounds by forming thioether adduct, Human embryonic stem cells (hES) derived MSC (hES-MSC) were FACS sorted as GSHhigh verus GSHlow fractions. The hES-MSCs in GSHhigh fraction stimulated chemoattraction capacity to platelet-derived growth factor (PDGF) and phosphorylation of both MAPK p42/44 and AKT which is involved in signaling transduction for stem cell migration. GSHhigh fraction cells also increased self renewal activity in colony forming unit of fibroblasts assay. As result of gene expression analysis, the hES-MSCs in GSHhigh fraction highly express the several genes including CXCR4, cMET, PDGF-RA, PDGF-RB, VEGF-R1, VEGF-R2, ANGPT1, and IDO2. Accordingly, GSHhigh fraction hES-MSCs significantly improved their therapeutic outcome to alleviate the airway inflammation in an experimental allergic asthma model. Taken together, these results demonstrates that intracellular level of GSH is novel indicator to determine the therapeutic efficacy of MSCs.