Graduation Year

2019

Document Type

Thesis

Degree

M.S.B.E.

Degree Name

MS in Biomedical Engineering (M.S.B.E.)

Degree Granting Department

Biomedical Engineering

Major Professor

Mark Jaroszeski, Ph.D.

Committee Member

Andrew Hoff, Ph.D.

Committee Member

Richard Gilbert, Ph.D.

Keywords

Atmospheric Plasma, Electroporation, Electrotransfer, Gene Delivery, Immunotherapy

Abstract

Gene therapy and immunotherapy are new up and coming fields in cancer treatment. In August of 2017, the first gene therapy was FDA approved called CAR-T cell therapy("FDA approval brings first gene therapy to the United States," 2017). This therapy uses the patient’s

own T-cells that are genetically modified to attack the cancerous cells. Currently, it takes 4-6 weeks to genetically modify these T-cells, which can be the difference between life and death for a cancer patient (Cheng, Jun, Jiang, & Xi, 2017). There is evidence that gene electrotransfer can help expedite the genetic modification process by having an easy to use device within the hospital. This could prevent the cells from having to be sent to a specialty lab, as is currently done, and would save shipping time. Traditionally, electroporation was a method that was considered for investigation; however, the main drawback is the need for an electrode to be in contact with the cells they are affecting. This led to the idea of using a newer electrogenetransfer method that involved using corona charge. Corona charge, also known as atmospheric plasma, has been defined as the gathering of charged particles in a neutral fluid (Chelsea M. Edelblute, Heller, Malik, & Heller, 2015; Ramachandran, Jaroszeski, & Hoff*, 2008). The advantage of corona charge is that it does not require electrodes to directly contact cells in order to have an electrical effect. In this research, the use of corona charge was investigated to determine if it caused an increase in molecular delivery across the membrane in a T-cell line compared to no treatment. The addition of heat at elevated temperatures 37°C, 40°C, and 43°C was determined if they caused a statistical increase alone and in combination with corona charge. Also, cell viability was also tested because the cells needed to maintain high cell viability to be used in treatment. T-tests were done to determine if the difference between treated and untreated cells was statistically significant. It was shown that treating the cells with corona charge for 3 minutes at 10kV and 25μA caused a statically significant increase in molecular delivery while maintaining high viability. Heat did not cause a statistically significant effect on molecular delivery. Combined corona charge treatment and heating to 37° and 40° C resulted in a statistically significant increase in molecular delivery compared to controls that were only heated. Additionally, combined corona charged treatment and heating to 40°C when compared to a control at room temperature, showed a statistically significant increase in molecular delivery in comparison to a sample that underwent corona charged treatment at room temperature.

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