Peripheral blood derived mononuclear cells (PBMCs) are important mediators involved in immune surveillance and are actively explored as the foundation of a wide range of therapies . It is convenient to isolate the cells from patients and re-administer them after desired manipulations. There are intensive efforts to genetically modify the cells using vectors that permanently integrate their cargo into cellular genome. Transient expression of transgenes, on the other hand, is more desirable to exert some control over the expression kinetics. Transient expression can be implemented with plasmid DNA (pDNA) or messenger RNA (mRNA) based expression systems . Non-viral delivery of expression systems minimizes any effects on the host genome and leads to a safer intervention. Transfection reagents developed by RJH Biosciences are particularly suitable for this application since they are highly compatible with human cells and achieve elevated levels of delivery. Lipofection has been alternatively explored for modification of primary cells and a number of leading commercial reagents are based on lipid based reagents. This application note summarizes experience with transfection of PBMC (and suspension growing Jurkat Cells) with pDNA and mRNA, comparing commercial lipofection and polymer reagents to a leading reagent developed by RJH Biosciences.
To assess efficiency of the transfection reagents in PBMC, we used pDNA and mRNA based expression systems for Green Fluorescent Protein (GFP). Both expression systems were obtained from commercial vendors and GFP expression was investigated after 2 days. Optimal ratios recommended by vendors were used to form the complexes, as well as an excess ratio beyond the recommended range. As shown in Figure 1, GFP expression was readily obtained with ALL-Fect using the pDNA based expression system, which was superior to the lipofection reagents used here.