A Transfection Device to Modify Mammalian Cells

Rudolph, Luke; Melvin, Jack; Sevilla, Donovan; Williams, Cassidy

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A Transfection Device to Modify Mammalian Cells

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Transfection is the introduction of macromolecules like nucleic acids and proteins into eukaryotic cells and has many applications in cellular modification for gene and cellular therapy. Efficient transfection of primary mammalian cells is crucial for gene and cellular therapy but remains a challenge. Viral vectors, like adenoviruses and lentiviruses, are effective but pose health risks to patients. Therefore, the development of a nonviral transfection technology to introduce nucleic acids and proteins into human primary cells was envisioned. The selected design was a microfluidic device that would use hydrodynamic shear forces generated as the cells pass through it. Cell viability was tested to determine the ideal parameters for cell safety. Proof of concept was tested using fluorescein-conjugated dextran sulfate, which demonstrated and confirmed the hypothesis that the shear forces were creating pores, allowing transfection to occur. DNA transfection was confirmed using a red fluorescent protein expressing plasmid. Protein transfection using a fluorescently labeled 22 kDa protein showed greater than 95% transfection efficiency. The results indicate successful transfection.

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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