Friday, October 28, 2011
Hall 1-2 (San Jose Convention Center)
Non-viral gene delivery is a promising field of study that has widespread applications in gene therapy and regenerative medicine. However, the low transfection efficiency of non-viral vectors remains a major limitation. To enhance the efficiency of non-viral gene transfer the physical (size and charge of the nanoparticles) and chemical (chemical structure of the polymers) properties of the vectors have been extensively studied while the cell and its microenvironment have not. A previous study in the Segura laboratory has shown that when cells were plated on fibronectin gene delivery was enhanced, whereas collagen I was shown to inhibit gene delivery in mouse mesenchymal stem cells (mMSCs). In the present study, mMSCs were cultured on surfaces coated with a combination of proteins such as fibronectin, collagen I, collagen IV, laminin, and vitronectin. Gene transfer was analyzed using a gaussia luciferase assay. To study cell morphology, actin and cell nucleus were stained using fluorescent immunohistochemistry. The degree to which gene transfer was enhanced or inhibited varied among groups, but those which included fibronectin, except when joined with collagen I, had an enhancement in gene transfer when compared to an uncoated surface. Coating was also shown to affect cell morphology, with cells plated on a combination including fibronectin resulting in more well-spread cells than those which contained collagen I. Our results indicate that the cellular microenvironment plays a role in non-viral gene transfer to mouse mesenchymal stem cells, and finding the optimal combination of ECM proteins can enhance non-viral gene transfer to these cells.