A transfection device to modify mammalian cells (SA2-2202)

Sandoval, Luis; Tweedie, Shelby; Bozza, Anthony; Conway, Walter

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A transfection device to modify mammalian cells (SA2-2202)

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Current methods of cellular transfection have several limitations, including low throughput, inefficiency, and low cell viability, as well as cytotoxicity. The major limitation is that with current methods, only one type of molecule can be transfected at once. There is a need for a device capable of transfecting a combination of DNA, RNA and/or proteins into mammalian cells with high throughput and efficiency. This project aimed to create a filtroporation-based device capable of deforming cell membranes to create transient pores allowing for macromolecule entry. The final design includes a closed system powered by a peristaltic pump with a 3D-printed interlocking device to support a series of membrane filters containing micropores that cells are forced through. This system demonstrated successful transfection of mammalian 3T3 cells with dextran sulfate of molecular weights 3-5 kDa and 20 kDa, a 22 kDa protein, as well as pmCherry2-N1 and pCMV-EYFP-NUC DNA, while achieving 95% cell viability. Ultimately, this system served as a successful proof of principle for this type of filtroporation approach to cellular transfection and alludes to the potential for this technology to revolutionize cell-based therapies and transgenic research.

This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

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