Non-viral and high-efficiency DNA delivery for transient nanog overexpression in mesenchymal stem cells by magnetofection
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Mesenchymal Stem Cells (MSCs) are a widely accepted and promising cell source for regenerative medicine. One major hindrance for practical use of MSCs is donor aging or cultural senescence. Previously, our group demonstrated that overexpression of Nanog can reverse the effects of organismal aging on MSCs proliferation and myogenic differentiation. Here, we tested the hypothesis that transient overexpression of Nanog in MSCs by non-viral DNA transfection may also promote proliferation and differentiation potential. We proposed to use Magnetofection (MF) as a non-viral method to overcome the low transfection efficiency in MSCs. Using human hair follicle derived MSCs (hHF-MSCs), gene transfer was first optimized with an EGFP-expressing plasmid. The optimized protocol was then applied to transfect hHF-MSCs with a Nanog-expressing plasmid. Overall, our results suggest that using MF can overcome the difficulty of DNA delivery into MSCs without significant cytotoxicity and increase efficiency greater than that of other widely used non-viral technologies.