Mentored Undergraduate Research Awards
Trehalose, a non-reducing disaccharide, is present in many microorganisms and metazoans. In these organisms, trehalose acts as a stress protectant and helps preserve lipid membranes of cells during states of desiccation and freezing. Trehalose is required on both sides of the cell membrane to achieve a significant cryoprotective effect. Specific loading methods for trehalose are required since this sugar is impermeant to mammalian cells. Trehalose loading in mammalian cells has been achieved by fluid-phase endocytosis and genetic modification for the expression of trehalose transporters, however cryoprotective outcomes are unable to compete with established methods of cryopreservation for mammalian cells. Sonoporation was achieved using a microfluidics device modified with an ultrasound emitter in the presence of microbubbles. Ultrasound frequencies emitted by the transducer result in a process called cavitation, which is the rapid expansion and collapse of lipid-coated gas-filled bubbles present in the solution. Cavitation of microbubbles creates small jets of liquid that can create membrane pores that are 150-300 nm in size and quickly reseal through budding and exocytosis allowing for uptake of impermeant compounds, such as trehalose.
Shaffer, Charles W. IV; Grimm, David F.; Menze, Michael A.; and Kopechek, Jonathan A., "Sonoporation-mediated loading of trehalose in cells for cryopreservation." (2020). Undergraduate Research Events. 14.