Non-viral gene delivery of DNA polyplexed with nanoparticles transfected into human mesenchymal stem cells.

Abstract

Human mesenchymal stem cells (hMSCs) represent a potent target for gene delivery for both stem cell differentiation applications and clinical therapies. However, it has, thus far, proven difficult to develop delivery vehicles that increase the efficiency of gene delivery to hMSCs, due to several problematic issues. We have evaluated different vehicles with regard to the efficiency with which they deliver hMSCs and enhance the ability to deliver a reporter gene. In this study, a non-viral gene delivery system using nanoparticles was designed, with emphasis placed on the ability of the system to mediate high levels of gene expression into stem cells. Via polyplexing with polyethylenimine (PEI), the cell-uptake ability of the nanoparticles was enhanced for both in vitro and in vivo culture systems. In experiments with PEI/pNDA polyplexed with nanoparticles, the expression of green fluorescent protein (GFP) with this vehicle was noted in up to 75% of hMSCs 2 days after transfection, and GFP gene expression was detected via Western blotting, flow cytometric analysis, and immunofluorescence using a confocal laser microscope after transfection.