Membrane permeabilization of mammalian cells using bursts of high magnetic field pulses

Abstract

Cell membrane permeabilization by pulsed electromagnetic fields (PEMF) is a novel contactless method which results in effects similar to conventional electroporation. The non-invasiveness of the methodology, independence from the biological object homogeneity and electrical conductance introduce high flexibility and potential applicability of the PEMF in biomedicine, food processing, and biotechnology. The inferior effectiveness of the PEMF permeabilization compared to standard electro-poration and the lack of clear description of the induced transmembrane transport are currently of major concern. Methods. The PEMF permeabilization experiments have been performed using a 5.5 T, 1.2 J pulse generator with a multilayer inductor as an applicator. We investigated the feasibility to increase membrane permeability of Chinese Hamster Ovary (CHO) cells using short microsecond (15 μs) pulse bursts (100 or 200 pulses) at low frequency (1 Hz) and high dB/dt (>106 T/s). The effectiveness of the treatment was evaluated by fluorescence microscopy and flow cytometry using two different fluorescent dyes: propidium iodide (PI) and YO-PROR© -1 (YP). The results were compared to conventional electroporation (single pulse, 1.2 kV/cm, 100 μs), i.e., positive control. Results. The proposed PEMF protocols (both for 100 and 200 pulses) resulted in increased number of permeable cells (70±11% for PI and 67±9% for YP). Both cell permeabilization assays also showed a significant (8±2% for PI and 35±14% for YP) increase in fluorescence intensity indicating membrane permeabilization. The survival was not affected. Discussion. The obtained results demonstrate the potential of PEMF as a contactless treatment for achieving reversible permeabilization of biological cells. Similar to electroporation, the PEMF permeabilization efficacy is influenced by pulse parameters in a dose-dependent manner.