Mesenchymal Stem Cells Engineering: Microcapsules-Assisted Gene Transfection and Magnetic Cell Separation

Stem cell engineering, the manipulation and functionalization, involving genetic modification of cells can significantly expand their application for cell therapy in humans; however, reliable, standardized, and cost-effective methods for cell manipulation are required. Here we explored the potential of magnetic multilayer capsules to serve as universal platform for non-viral gene transfer, stem cell magnetization and magnetic cell separation to improve gene transfer efficiency. In particular the following experiments were performed: (i) the study of internalization process of magnetic capsules into stem cells, including capsule co-localization with established markers of endo-lysosomal pathway,(ii) characterization and quantification of capsule uptake with confocal, electron microscopy and flow cytometry, (iii) intracellular delivery of messenger RNA (mRNA) and separation of gene-introduced cells by MACS procedure, (iv) the influence of capsules on cell proliferation potential. Importantly, due to interna...