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ETM study of electroporation influence on cell morphology in human malignant melanoma and human primary gingival fibroblast cells
Authors:Nina Skolucka  Malgorzata Daczewska  Jolanta Saczko  Agnieszka Chwilkowska  Anna Choromanska  Malgorzata Kotulska  Iwona Kaminska  Julita Kulbacka
Institution:1.Department of Medical Biochemistry, Medical University, Chalubinskiego 10 St., 50-368 Wroclaw, Poland;2.Department of General Zoology, Zoological Institute, University of Wroclaw, Sienkiewicza 21 St., 50-335 Wroclaw, Poland;3.Institute of Biomedical Engineering and Instrumentation, Wroclaw University of Technology, ul. Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland
Abstract:

Objective

To estimate electroporation (EP) influence on malignant and normal cells.

Methods

Two cell lines including human malignant melanoma (Me-45) and normal human gingival fibroblast (HGFs) were used. EP parameters were the following: 250, 1 000, 1 750, 2 500 V/cm; 50 µs by 5 impulses for every case. The viability of cells after EP was estimated by MTT assay. The ultrastructural analysis was observed by transmission electron microscope (Zeiss EM 900).

Results

In the current study we observed the intracellular effect following EP on Me-45 and HGF cells. At the conditions applied, we did not observe any significant damage of mitochondrial activity in both cell lines treated by EP. Conversely, we showed that EP in some conditions can stimulate cells to proliferation. Some changes induced by EP were only visible in electron microscopy. In fibroblast cells we observed significant changes in lower parameters of EP (250 and 1 000 V/cm). After applying higher electric field intensities (2 500 V/cm) we detected many vacuoles, myelin-like bodies and swallowed endoplasmic reticulum. In melanoma cells such strong pathological modifications after EP were not observed, in comparison with control cells. The ultrastructure of both treated cell lines was changed according to the applied parameters of EP.

Conclusions

We can claim that EP conditions are cell line dependent. In terms of the intracellular morphology, human fibroblasts are more sensitive to electric field as compared with melanoma cells. Optimal conditions should be determined for each cell line. Summarizing our study, we can conclude that EP is not an invasive method for human normal and malignant cells. This technique can be safely applied in chemotherapy for delivering drugs into tumor cells.
Keywords:Electroporation  Electron microscopy  Human melanoma cells  Primary fibroblasts  Cell morphology  Transmission electron microscope  Malignant cell  Normal cell  Intracellular effect  Electric field  Proliferation  Ultrastructural analysis
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