Josiah Rudge

BioE Ph.D. Defense Presentation

Time and Date: 3:00 p.m., Wednesday, April 9th, 2025

Location: EBB CHOA

Advisor:

Aniruddh Sarkar, Ph.D. (Georgia Institute of Technology)

Commitee:

John Blazeck, Ph.D. (Georgia Institute of Technology)

Gabe Kwong, Ph.D. (Georgia Institute of Technology)

David Myers, Ph.D. (Georgia Institute of Technology)

Fatih Sarioglu, Ph.D. (Georgia Institute of Technology)

Single-Cell Impedance Cytometry Enabling Novel Single-Cell Electroporation Capabilities in a High Throughput Device

Cell therapies have shown great promise in treating diseases such as CAR-T cells for blood cancers. Manufacturing these therapies presents considerable challenges and costs. These include quality of donor source material, transfection of cells, and controlling and measuring the quality of the product. Current therapies often transfect cells by viral vectors which are costly, have payload limitations, are difficult to target specific cells with, and present safety concerns due to immunogenicity and oncogenicity. A microfluidic single-cell electroporation device and scheme was made to address these concerns. Electroporation is an alternative non-viral method of transfection that is inexpensive and non-immunogenic. Additionally, it is applicable to different cell types or delivery payloads including larger payloads (e.g. CRISPR-Cas). However, electroporation can cause significant cell death, toxicity, and/or low delivery efficiencies. This microfluidic device electronically measures properties of single cells and applies an electroporation voltage based on that measurement. This feedback is demonstrated by selectively delivering to only certain cell types in a mixture of cell types. Further efforts were made to create a PDMS-on-glass version of the device to be compatible with microscopy imaging, as well as producing functional CAR-T cells with the device.