Abstract 2438 Enhancing Lipofectamine Transfection Efficiency using Capillary-Mediated Vitrification Process

Transfecting primary mammalian cells presents challenges in achieving effective nucleic acid introduction. Lipofectamine, a widely used transfection reagent, employs a reputable lipid-based delivery method. However, difficulties persist, particularly in achieving effective mRNA transfection in challenging cell lines. Addressing this issue, we developed a technology aimed at preserving biomolecules, including mRNA, against ambient stressors without compromising its structural and functional integrity. Focusing on Lipofectamine MessengerMax as a cargo delivery system, we utilized mRNA encoding eGFP to assess transfection efficiency in Chinese Hamster Ovary (CHO) cells. Employing our novel vitrification technology and Biofix™ Buffer, we aimed to stabilize these transfection reagents on a porous scaffold for long-term storage and downstream application in cells. Our results indicate that both lipofectamine and mRNA are highly compatible with our vitrification process and buffer. Additionally, the use of stabilized lipofectamine reagent led to a greater than two-fold increase in transfection relative to the standard transfection reagents. Preliminary data also demonstrates consistently enhanced transgene expression even after storing lipofectamine at room temperature and 37°C for a month on our scaffold. These results showcase the potential of our innovative approach to enhance transfection while maintaining the functionality of the reagents.