Comparison of efficiency and toxicity of transfection reagents carrying different amounts of DNA for mesenchymal stem cells (MSCs). These cells are primary cells and considered hard-to-transfect. Panel A shows the transfection efficiency of “Rutgers Transfection System” in comparison to the five most widely used commercially available reagents; Panel B shows the viability of cells after transfection with “Rutgers Transfection System” in comparison to the same five commercially available ones. The most efficient vectors and their corresponding toxicities are highlighted with red arrows.
Invention Summary: For various biomedical research and preclinical or clinical applications, stem cells (SCs) are transfected ex-vivo with transgenes to either express a biomolecule or differentiate in a certain way. This genetic engineering process usually involves the use of a gene delivery system (vector) to transfect the SCs. Many research laboratories prefer to use non-viral vectors to genetically modify SCs because they are more cost-effective and, in general, safer than viral vectors. However, commercially available non-viral vectors (polymers and lipids) not only have low transfection efficiency (at best 30%), but also could negatively impact SC viability and/or cause genotoxicity. Electroporation is another method for SC transfection, but it leads to excessive cell death (>60% toxicity). Therefore, there is a need for the development of a vector that is both efficient and safe. Rutgers Scientists have developed a gene transfer system that include a novel recombinant non-viral fusion peptide vector and a transfection cocktail that have been optimized to achieve high transfection efficiency without significant somatic- or genotoxicity. Side-by-side comparison with five commercially available transfection reagents that are advertised for SC transfection demonstrated that our system leads to more than 50% efficiency, while the commercially available reagents only had 30% or less efficiency. Further, the scientists investigated cell proliferation rates, membrane integrity and morphology, genotoxicity, gene regulation (microarray analysis), SC surface biomarkers and SC differentiation. In all these studies, minimal negative impact on SCs’ gene function, viability or ability to differentiate was observed with the Rutgers transfection system. For detailed information, please see recently published on article on Pubmed (PMID: 29078136) or at: https://doi.org/10.1016/j.biomaterials.2017.10.028. Market Applications:
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Intellectual Property & Development: Patent pending. Available for licensing and/or research collaboration.