Abstract: Transfection of nucleic acids into blood cells holds great potential for various therapeutic applications. Lipid-based transfection reagents have emerged as a promising approach for delivering nucleic acids into cells due to their versatility and biocompatibility. In this study, we aimed to compare and evaluate the efficiency of different lipid-based transfection reagents for delivering nucleic acids into blood cells.
Methods: Blood samples were collected from healthy human volunteers, and peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation. We tested five different commercially available lipid-based transfection reagents: Reagent A, Reagent B, Reagent C, Reagent D, and Reagent E. Each reagent was used to transfect PBMCs with a plasmid DNA encoding a fluorescent reporter gene. Transfection efficiency was assessed by flow cytometry, measuring the percentage of fluorescent cells.
Results: Our results demonstrated that all five lipid-based transfection reagents were capable of delivering nucleic acids into PBMCs. However, significant variations in transfection efficiency were observed among the different reagents. Reagent C consistently showed the highest transfection efficiency, resulting in an average of 60% fluorescent cells. Reagent A and Reagent E exhibited moderate transfection efficiency, with approximately 40% and 35% fluorescent cells, respectively. Reagent B and Reagent D yielded the lowest transfection efficiency, with less than 20% fluorescent cells.
Discussion: The variations in transfection efficiency among the tested lipid-based reagents could be attributed to differences in their formulation, chemical properties, and interactions with blood cells. Lipid-based reagents facilitate the formation of stable complexes with nucleic acids, promoting their uptake by cells. The observed differences in transfection efficiency highlight the importance of selecting the appropriate lipid-based reagent for specific blood cell types and desired transfection outcomes.
Conclusion: Our study demonstrates that lipid-based transfection reagents can efficiently deliver nucleic acids into blood cells, with varying levels of transfection efficiency. Reagent C exhibited the highest efficiency, making it a promising candidate for blood cell transfection applications. The findings of this study provide valuable insights for researchers and clinicians seeking to optimize transfection protocols for manipulating blood cells using lipid-based reagents.