PEDOT:PSS Enhanced Electroporation for Cell Therapy Manufacturing

Cellular immunotherapies, such as chimeric antigen receptor (CAR) T cell therapies and macrophage therapies, have shown promising results in treating a wide range of cancers. Advances in these therapies focus on increasingly complex cellular systems to simultaneously optimize immune cell trafficking to the tumor microenvironment (TME), TME control, tumor recognition, healthy tissue discrimination, proliferation, persistence, and external feedback control. The typical production of these therapies incorporates various conventional cell engineering strategies, such as viral vectors, lipid vectors, electroporation, microinjection, and chemical transfection, among others. Miniaturization and automation of these techniques are essential for scaling the complexity of cellular therapies without concurrently scaling the costs of research and production. To achieve this miniaturization, recent advances in microelectroporation, organic electronics, and microfluidics can be applied to create an integrated platform to efficiently fabricate and interrogate arbitrarily complex cellular therapies. Myeloid cells in particular, such as macrophages, have been a recent focus for novel cellular therapies due to their unique tissue infiltration capabilities and innate immune effector functions that allow them to overcome challenges in solid tumor infiltration. Despite their promise, efficiently modifying myeloid cells remains a major challenge because of their limited expansion capacity and particular sensitivity to transfection and transduction processes. Although pulsed electric field electroporation effectively permeabilizes cell membranes to deliver payloads without the need for toxic chemical or viral transduction agents, conventional bulk electroporation devices face major challenges with cell viability, heterogeneity, and electrochemistry at the electrode-electrolyte interface. These challenges inspired this work, which introduces a versatile platform for efficient transfection and transduction of cells with therapeutic potential. This platform demonstrates the use of polymer electrodes for cell positioning and stimulation, which substantially increases cell viability and transfection efficiency. As a proof-of-concept, we demonstrate enhanced delivery of Cas9 protein, gRNA, and plasmid DNA into cell lines and primary cells. We further characterized the therapeutic efficacy of modified macrophages by modulating their polarization state in both in vitro and in vivo tumor models. These findings demonstrate a novel approach for rapid modification of difficult-to-transfect cell types to accelerate their study and use as therapeutic platforms.

Name dc:type.qualificationname

Doctor of Philosophy (PhD)

Level dc:type.qualificationlevel

Doctoral

Grantor dc:publisher.institution

University of Cambridge

Year dc:date.issued

2025

Author dc:creator

• Gharia, Asmaysinh

Advisors dc:contributor.advisor

• Malliaras, George
• Fraser, Iain

• bioelectronics
• microfluidics
• electroporation
• cell therapy
• organic electronics

Licence

• https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/3477d875-e273-46f8-8a4c-a9623c476dca/download
• http://purl.org/NET/rdflicense/allrightsreserved

DOI dc:identifier.doi

https://doi.org/10.17863/CAM.119715

OAI identifier oai:identifier

oai:www.repository.cam.ac.uk:1810/386555