Abstract
Intracellular delivery of DNA is considered a challenge in biological research and treatment of diseases. The previously reported transfection rate by commercially available transfection reagents in cancer cell lines, such as the mouse lung tumor cell line (TC-1), is very low. The purpose of this study is to introduce and optimize an efficient gene transfection method by mechanical approaches. The combinatory transfection effect of mechanical treatments and conventional chemical carriers is also investigated on a formerly reported hard-to-transfect cell line (TC-1). To study the effect of mechanical loadings on transfection rate, TC-1 tumor cells are subjected to uniaxial cyclic stretch, equiaxial cyclic stretch, and shear stress. The TurboFect transfection reagent is exerted for chemical transfection purposes. The pEGFP-N1 vector encoding the green fluorescent protein (GFP) expression is utilized to determine gene delivery into the cells. The results show a significant DNA delivery rate (by ~30%) in mechanically transfected cells compared to the samples that were transfected with chemical carriers. Moreover, the simultaneous treatment of TC-1 tumor cells with chemical carriers and mechanical loadings significantly increases the gene transfection rate up to ~ 63% after 24 h post-transfection. Our results suggest that the simultaneous use of mechanical loading and chemical reagent can be a promising approach in delivering cargoes into cells with low transfection potentials and lead to efficient cancer treatments.
Highlights
Managing the transportation of molecules in biological cells is a significant aim in many medical processes including gene therapy and treatment of diseases such as cancer and viral diseases [1]
This paper investigated the transfection efficiency of pEGFP-N1 in TC1 cell line
Shahbazi et al [43] showed the pEGFP-N1 and pEGFP-E7 delivery were detected in approximately 99.1% and 80.63% of HEK-293T cells, 5.04% and 4.47% of TC-1 cells, and 6.66% and 5.95% of A549 cells treated with TurboFect, respectively
Summary
Managing the transportation of molecules in biological cells is a significant aim in many medical processes including gene therapy and treatment of diseases such as cancer and viral diseases [1]. From the various DNA transfection methods applied for eukaryotic cells, some methods rely on physical treatments and others rely on chemical materials or biological particles as the functional carriers. Chemical carrier can have a polymeric (such as polyplexes) or a lipid base (such as lipofection) [1]. Transfection using physical methods, such as electroporation and sonoporation, is challenging as they showed to physically disrupt the cell membrane [2]. Many researchers have developed several physical methods for gene transfection [3,4,5]
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