Genetic engineering of stem cells by non-viral vectors

Stem/progenitor cells hold a great promise for application in several therapies due to their unique biological characteristics. With the purpose of harnessing these cells full potential in cell-or gene-based therapies it might be advantageous to enhance some of their features through gene delivery strategies. Accordingly, we are interested in developing efficient and safe methodologies to genetically engineer stem cells, boosting their therapeutic efficacy in Regenerative Medicine. In our work, delivery of plasmid DNA to human Bone Marrow Mesenchymal Stem Cells (BM-MSC) was optimized by lipofection and by a recently available microporation technique and no effect was observed in their immunophenotypic characteristics or differentiative potential. After lipofection similar number of plasmid copies was determined at different cell passages. Importantly, cell proliferation kinetics slowed down due to the presence of plasmid. Overall, we believe our findings are extremely useful towards the maximization of gene delivery to human MSC, without compromising cell function and viability.