Effects Of Electromagnetic Pulse Field On Cell Electroporation

The effects of electromagnetic pulse field on cell electroporation were studied in this research. In the first part, the mixture of full yeast cells (54.S.cerevisiae DC5 (a, leu2-3, 1eu2-112, his-39, canl-11) or their protoplasm and plasmid (YRP12-54) were treated in low electromagnetic pulse field provided by Broad Band Transverse Electromagnetic Cell (BTEM Cell). The results indicated that plasmids could enter the full yeast cells when they were treated for 1, 5, 10, 20 and 40 min and the transformation rates were 4.6 ×10², 5.2 ×10², 1.46 ×10³, 2.78 × 10³ and 1.0×10³/μg DNA, respectively. The transformation rates for protoplasm were 6.6× 10², 4.88× 10², 2.16×10³, 5.24×10³ and 1.24×10³/μ.g DNA, respectively, when they were treated for 1, 5, 10, 20 and 40 min.The effects of high electromagnetic pulse field on electroporation of rabbit red blood cells were also studied. The Gene Pulser Transfection Apparatus was used to generate high electromagnetic pulse field. Holes with diameter 0.5-2 μm on the treated cell membrane were observed under a scan electro-microscope. In the experiments of Trypan blue stain, we used 4 pulses, 1000 volt, and capacitance of 4 μp, 6 μF, 8 μF, 10 μF, the reversible electroporation percentage was 17.4%, 26.8%, 31.6%, 29.0%, respectively. In another experiment, 4 pulses, 8 μF capacitance and voltages of 300, 600, 1000 volt were used and the reversible electroporation percentage were 14.0%, 24.2%, 30.8%, respectively.The cell toxicity of cyclophosphamide was also studied in high electromagnetic pulse field generated by Gene Pulser Transfection Apparatus. In the cyclophosphamide concentration of 0.03 μg/mL, 0.3 μ.g/mL, 3μg /mLwithout electromagnetic pulse field, the chromosome aberrance percentages were 7.3%, 12.0%, 18.3%, respectively, compared to 12.3%, 18.3%, 32.3% in high electromagnetic pulse field. This result indicated that high electromagnetic pulse field has significant effect on the cell toxicity of cyclophosphamide.