(208a) Integration of Electroporation-Based Gene Transfection and Nanofiber-Based Cell Culture for Embryonic Stem Cells | AIChE

(208a) Integration of Electroporation-Based Gene Transfection and Nanofiber-Based Cell Culture for Embryonic Stem Cells

Authors 

Fei, Z. - Presenter, The Ohio State University
Sharma, S. - Presenter, The Ohio State Univeristy
Lannutti, J. J. - Presenter, The Ohio State University
Lee, L. J. - Presenter, The Ohio State University


Over the past decade, the use of genetically modified embryonic stem (ES) cells and iPS cells has gained prominence as an attractive tool for a wide range of clinical applications. So far, this area is dominated by viral-mediated transduction, which is efficient but safety issues have hampered their clinical uses in humans. Non-viral methods are advancing as promising alternate approaches. However, low delivery efficiency of the therapeutic nucleic acid into the nucleus of the target cell is a significant obstacle in non-viral gene therapy. Previously, we demonstrated an electroporation-based gene delivery method, called membrane sandwich electroporation (MSE). The MSE method is able to provide better gene confinement near the cell surface to facilitate gene transport into the cells and thus shows significant improvement of transgene expression for mammalian cells over current electroporation techniques. Here, we report a novel method that integrates MSE for gene delivery to embryonic stem cells (ESC) cultured on electrospun gelatin/PCL nanofibers. Electrospun nanofibers are known to provide an in vivo-like micro-environment for cell culture. It has great potential to replace the widely used matrigel, which is reduced from bacteria, for safe ES cell culture. A combination of MSE and nanofibers allows for the cells to be cultured on the same substrate before and after gene transfection, thus reducing the number of harsh instances occurred in conventional electroporation, such as repeated cell trypsinization for getting single cell suspension, high voltage, and longer time duration outside the incubator. These advantages are particularly important for culturing delicate and hard-to-transfect primary human cells.