Strategy for creating biomedical cell-based products based on genome editing via plasmonic laser transfection

Objective: adaptation of genome reprogramming technology based on laser transfection to create biomedical cell products. Material and methods. We developed optotransfection platforms based on multi-well culture plates with the working surface coated with gold nanoparticle monolayers. Model cell experiments on the delivery of the molecular agent propidium iodide were conducted using the mice macrophage cells, line RAW 264.7. Transfection of cell monolayers adhered to 24-well plate platforms with gold nanostar layers was carried out via single 2-D scanning with a pulsed laser and narrowly focused beam. The viability and efficiency of cell modification were assessed by direct microscopy and metabolic tests. Results. The optimal operating modes for laser optotransfection of RAW 264.7 mouse macrophage cells were determined, namely: pulse energy 1.4 μJ, pulse duration 200 ns, pulse frequency 10 kHz, scanning speed 0.025 m/s. The minimum starting monolayer confluency was 40%, the transfection efficiency was 78±7.6%, and the cell viability was 84±9.3%. Conclusion. Under controlled model in vitro experiments, it has been demonstrated that fine adjustment of optotransfection parameters enables reproducible results of genetic modification in lymphocytelike cells, achieving high levels of both efficiency and viability.