Abstract

Inserting foreign biomolecules into human cells is at the heart of cell engineering protocols. By taking nucleic acids or fully formed proteins and shuttling them across the cell membrane in a
process known as transfection, cells can either temporarily or permanently gain or lose functionalities. This capability has been used extensively for applications including fundamental
research into genetics, industrial production of high value biomolecules, and of interest to this thesis, the production of novel cell therapies – where human cells are repurposed to fight disease.
Numerous techniques have been developed to perform transfection on human cells with a specific focus on technologies that can engineer enough cells for clinical use (often > 10^9 cells are needed to treat a single patient). However, a currently unmet need in this field is a miniaturized platform for the research and development of new cell therapies. For this application, large
libraries of cellular modifications need to be tested in hopes of discovering one with clinical potential. To do this economically, testing each modification in the library must be done rapidly
while consuming as few resources as possible. Bulk microfluidics has emerged as an ideal technology for high throughput clinical cell therapy production; however, it is unsuited to processing numerous unique small-scale reactions in parallel. To address the unmet need, in this thesis we demonstrate that droplet microfluidics – the science of controllably manipulating sub-microliter volumes of liquid – can serve as the ideal platform for cell therapy R&D.