The art of delivery: uptake into living cells by microwave field induced perturbation of the plasma membrane

• Applicants: Professorin Dr. Maria Cristina Cardoso; Professor Dr.-Ing. Rolf Jakoby
• Subject Area: Electronic Semiconductors, Components and Circuits, Integrated Systems, Sensor Technology, Theoretical Electrical Engineering ; Cell Biology
• Term: from 2020 to 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 431063518

• Final Report Year: 2024

Final Report Abstract

The availability of powerful techniques that allow the non-invasive introduction of a variety of compounds to artificially manipulate cell function is essential for basic research and potential therapeutic applications. The cell membrane is the largest barrier for the transport of molecules into living cells. Therefore, there is great interest in effective cell transfection methods. Importantly, physical transfection methods enable the transport of molecules of different chemical compositions and are therefore very flexible. The development of new physical technologies to temporarily disrupt this barrier and enable the transport and incorporation of synthetic compounds into the molecular machinery of cells requires an interdisciplinary approach with expertise in engineering and biology. The recently discovered microwave-based transfection process offers the prospect of overcoming the problems of established state-ofthe-art physical transfection techniques. Based on our proof-of-concept results, in which we built and tested a prototype for microwave-assisted transfection that enabled the uptake of fluorescently labeled peptides into living cells, we first investigated the influence of electromagnetic radiation in the microwave range (GHz) of different intensity and frequency as well as the temperature influence on the plasma membrane of the cells, and thus on the transfection rate, and extended the previously obtained results to the uptake of a variety of molecules of different chemical composition and a wide size range into living cells. We also tested the effects of the newly developed technique on cell physiology and used it to study cell physiology in real time in living cells. In addition, the settling and firm growth of the cells as well as the dielectric differences between transfected and non-transfected cells were measured. Overall, the use of microwave radiation in the GHz range, especially at a frequency of 18 GHz, enables the uptake of various substances directly into mammalian cells growing as adherent cultures with little physiological impact and the choice of frequency plays an elementary role.

Publications

• Broadband Microwave Electroporation Device for the Analysis of the Influence of Frequency, Temperature and Electrical Field Strength. 2022 44th Annual International Conference of the IEEE Engineering in Medicine & Biology Society (EMBC), 1565-1568. IEEE.
  Paravicini, Markus; Milden, Manuela; Frank, Laura M. Pimentel Paes; Schussler, Martin; Cardoso, M. Cristina; Jakoby, Rolf & Hessinger, Carolin
  
• Sensing of Muscular Mouse Cells C2C12 from Seed Out to Electroporation - A Conceptional Study. 2022 IEEE/MTT-S International Microwave Symposium - IMS 2022, 802-805. IEEE.
  Paravicini, Markus; Milden, Manuela; Birnstengel, Daniel; SchuBler, Martin; Jakoby, Rolf; Cardoso, M. Cristina & Hessinger, Carolin
  
• Uptake of substances into living mammalian cells by microwave induced perturbation of the plasma membrane. Scientific Reports, 14(1).
  Milden-Appel, Manuela; Paravicini, Markus; Milden, Jannick P.; Schüßler, Martin; Jakoby, Rolf & Cardoso, M. Cristina