Influence of Submicro-Grooved Surface and Zwitterionic Molecule-Modified PEI on Gene Transfection
Date Issued
2014
Date
2014
Author(s)
Lian, Yen-Shiang
Abstract
Gene therapy is the use of molecular biology to treat disease by delivering therapeutic DNA into cells, which is well application and discussion on cancer, immune disease, tissue engineering and gene defects. Previous research has less discussion on cell morphology and gene transfection. In this work, we used different sub-micro polystyrene (PS) surface to observe the morphology of mouse myoblast cell C2C12, expecting to affect gene transfection efficiency by varying topography. We found that the transfection efficiency to elongated cells on the grooved surface was lower compared to the efficiency of the cells on the flat surface. The results indicate that nuclei morphology might affect the gene transfection by non-viral cationic polymeric vehicles. Then, we used zwitterionic molecule to modify Polyethylenimine (PEI) which was used as a carrier for gene transfection, hoping to decrease cell toxicity and increase gene transfection efficiency.
First, C2C12 cells were seeded on grooved polystyrene. PEI and DNA complexes for were applied on gene transfection. Lysing cells and quantified the fluorescence intensity of fluorescent Proteins to define gene transfection efficiency. The results indicated that the gene transfection efficiency of the cells on the grooved surface is less than which on the flat surface, and the trend became more obviously as ridge and width increased. Then, we will discuss the influence of different submicron surface topography on nucleus morphology and non-viral gene transfection. The results showed that nucleus deformed on the grooved surfaces, and the deformation became larger on the grooved surfaces. We also suggested that the arrangement of cytoskeleton was the important factor of gene transfection.
In addition, the zwitterionic molecule CBAAester was used to modify PEI, the cell viability was decrease and the gene transfection efficiency was enhanced. The grafting ratio of 6% has better gene transfection efficiency than other grafting ratios.
Subjects
基因轉殖
細胞型態
次微米地形
兩性離子分子
SDGs
Type
thesis
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