Optimization of Pulse Duration Parameter for Hela Cells Growth Rate

To introduce a cell or tissue with a gene or DNA, electroporation parameter plays the most important role. Research on electroporation parameters is still in its early stages. Different researches have used different parameters while performing their research. Electroporation is a mechanism of temporarily disrupting the bilayer membrane, in such a way that a hydrophilic pore is established, allowing a pathway into the cell for molecules such as DNA, which then heals up, once again protecting the cell from the outside. There are many theories as to how this can occur, but the simplest view is that a short pulse creates an increase in the trans-membrane potential which if it exceeds a certain threshold (dependent on size and shape of cells) can lead to a thinning of the bilayer, either due to a local dislocation in the membrane. In this study, cervical cancer cells (HeLa cells) was used to expose to single pulse electric field. Thus, for the purpose of this study, the field strength of 1kV/cm was selected and the pulse duration was varied (30µs, 70µs, 100µs, 200µs, 300µs and 600µs) to obtain higher proliferation for a growth rate of HeLa cells. From this study, it is determined that, HeLa cell exposed to 1kV/cm with a pulse duration of 100µs and single pulse revealed the highest and fastest percentage confluence when compared to growth rate of HeLa cell exposed for 30µs, 70µs, 200µs, 300µs and 600µs pulse duration

Investigation of Pulse electric field effect on HeLa cells alignment properties on extracellular matrix protein patterned surface

YesCell behavior in terms of adhesion, orientation and guidance, on extracellular matrix (ECM) molecules including collagen, fibronectin and laminin can be examined using micro contact printing (MCP). These cell adhesion proteins can direct cellular adhesion, migration, differentiation and network formation in-vitro. This study investigates the effect of microcontact printed ECM protein, namely fibronectin, on alignment and morphology of HeLa cells cultured in-vitro. Fibronectin was stamped on plain glass cover slips to create patterns of 25μm, 50μm and 100μm width. However, HeLa cells seeded on 50μm induced the best alignment on fibronectin pattern (7.66° ±1.55SD). As a consequence of this, 50μm wide fibronectin pattern was used to see how fibronectin induced cell guidance of HeLa cells was influenced by 100μs and single pulse electric fields (PEF) of 1kV/cm. The results indicates that cells aligned more under pulse electric field exposure (2.33° ±1.52SD) on fibronectin pattern substrate. Thus, PEF usage on biological cells would appear to enhance cell surface attachment and cell guidance. Understanding this further may have applications in enhancing tissue graft generation and potentially wound repair.Ministry of Higher Education Malaysia and UTHM Tier 1 Research Grant (U865