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Cell Electroporation with new Platinum Compounds

STSM by Elisabetta Sieni, Early-Stage Researcher

Period of mission: from 19/03/2015 to 30/03/2015

Home institution: Padova University Department of Industrial Engineering, Padova, Italy

Host institution: University of Ljubljana, Faculty of Electrical Engineering, Ljubljana, Slovenia

The idea was to use two new platinum compounds with different solubility in the culture media and study their behavior under electroporation conditions.

The cytotoxicity of two platinum compounds (named C, cis-guanidine and D, trans-guanidine) has been tested on B16F1 cells in two different conditions: without and with electroporation. The cytotoxicity without electroporation has been compared with the one of cis-Platin. 

Cytotoxicity without electroporation has been performed in 6 repetitions with different concentration of the compounds in order to evaluate the IC50. Cells have been plated and incubated 24h before adding compounds. The exposure to the compound has been performed for 5 min and 48h. The permeabilization as well as the voltage pulses survival for B16F1 cells has been verified for 6 different voltages applied between electrodes in the range 0 and 320 V using electrodes with 2 plates electrodes at 2mm between them and 8 pulses at 1 Hz (pulse length 100 µs). Permeabilization has been verified using Propidium Iodide (PI) fluorescent indicator and TECAN. From survival and permeabilisation results the suitable voltage intensity for this type of cells has been selected to be 200 V. Finally, cytotoxicity of compounds with and without electroporation has been verified by means of MTS test 72hafter the electroporation/exposition to Platinum compoundof cells seeding. It appears that the compound D shows a reduction of cell survival for a 5 min compound-exposition if voltage pulses have been applied. This difference was not observedin the experiments where the compound C was tested.


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