Investigation of Pulse Electric Field Influence on Cell Trypsinization Assay
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Keywords:
Attachment, pulse electric field, trypsinization, wound healing.Abstract
This study is aimed at investigating the effect of pulse electric field on cell attachment properties towards the enhancement of wound healing process. The Human Colorectal Adenocarcinoma cell line (HT29) was use in this study. The HT29 cells were treated with an electric field of 600V/cm amplitude for 500µs pulse duration in suspension and seeded for 24 hours. A time-lapse live imaging of the trypsinization assay of the HT29 cell was carried out using integrated devices for live cell imaging that was equipped with Charge Coupled Device (CCD) Camera, Temperature and Carbon IV Oxide (CO2) Controllers as well as, an Inverted Microscope. The electric treatment was found to decrease the adhesiveness of the cell by 41% where the treated cells detached from the substrate in 340 seconds as compared to control group that took 480 seconds to completely detached from the substrate immediately after trypsinization. Hence, the study suggested that the application of appropriate electric field, can affect the cell signalling pathways which in turn decrease the degree of cell adhesion on the substrate. The decrement in the cell adhesion could facilitate wound healing process via increased cellular migration, since the speed of cell migration is inversely proportional to the strength of cell adhesion.
References
Pehlivanova VN, Tsoneva JH & Tzoneva RD (2012), Multiple ef-fects of electroporation on the adhesive behavior of breast cancer cells and fibroblast. Cancer International, 12(9), 1-9
Hondroulis E, Melnick SJ, Zhang X, Wu ZZ & Li CZ (2013), Elec-tric field manipulation of cancer cell behavior monitored by whole-cell biosensing device. Biomed microdevices, 15, 657-663
Gekas J, Hindié M, Faucheux N, Lanvin O, Mazière C, Fuentès V & Nagel MD (2004), The inhibition of cell spreading on a cellulose substrate (cuprophan) induces an apoptotic process via a mitochon-dria‐dependent pathway. FEBS Letters, 563(1-3), pp. 103-107.
Kotnik T, Kramar P, Puchir G & Miklavcic D (2012), Cell Mem-brane Electroporation Part1: Phenomenon. IEEE electrical insula-tion magazine, 28 (5), 14-23
Kotnik T, Pucihar G & Miklavcic D (2010), Induced transmem-brane voltage and its correlation with electroporation-mediated mo-lecular transport. The Journal of Membrane Biology, 236(1), 3–13
View more references (9)
Serša G, Čemažar M, Miklavčič D, Rudolf Z (2006), Electrochemo-therapy of tumors. Radiology & Oncology, 40, 163–174
Sersa G, Kranjc S, Scancar J, Krzan M & Cemaza M (2010), Elec-trochemotherapy of Mouse Sarcoma tumors using electric pulse trains with repetition frequencies of 1Hz and 5 kHz. J Membrane Biol, 236(1), 155-162
Mir LM, Gehl J, Sersa G, Collins CG, Garbay JR, Billard V & Marty M (2006), Standard operating procedures of the elec-trochemotherapy. European Journal of Cancer Supplements, 4(11), 14–25
Gaynor P, Wells DN & Oback BB (2005), Couplet alignment and improved electrofusion by dielectrophoresis for a zona-free high-throughput cloned embryo production system. Medical and Biologi-cal Engineering and Computing, 43, 150–154
Lye H (2012), Growth Properties and Cholesterol Removal Ability of Electroporated Lactobacillus acidophilus BT 1088. Journal of Microbiology and Biotechnology. 22 (7), 981-989
Rodamporn S (2011), Optimal parameters of electroporation for gene and tissue. Biomedical Engineering International Conference. 279-282
Rubinsky B (2007), Irreversible electroporation in medicine. Tech-nology in Cancer Research & Treatment, 6(4), 255–260
Lodish,H, Berk A, Zipursky SL, Matsudaira P, Baltimore D & Dar-nel J (2000), Molecular cell biology (4th ed.). New York: W. H. Freeman
Sevilla C (2012), The role of extracellular matrix fibronectin and collagen in cell proliferation and cellular self-assembly. University of Rochester, UK. Ph.D. thesis