Aranda Hernández, Jorge; (2025) Microfluidic Devices as Tools for Cell Therapy Manufacturing. Doctoral thesis (Eng.D), UCL (University College London).

Text Aranda Hernandez_10209079_thesis.pdf Access restricted to UCL open access staff until 1 June 2026. Download (4MB)

Abstract

The advent of cellular adoptive immunotherapy, such as chimeric antigen receptor (CAR) T cell therapies is a paradigm shift for the field of personalised medicine. CAR T cells have immense potential as an effective treatment for blood cancers. A key bottleneck for the clinical translation of CAR T cell therapies is the expensive and lengthy manufacturing process, particularly during the gene delivery and expansion steps. Automated and functionally closed, bioprocessing systems, also known as GMP-in-a-box systems, have emerged to tackle these challenges. Using integrated sensors, single-use vessels and process automation, these systems have been shown to increase the efficiency of CAR T cell manufacturing processes and have the potential to reduce the costs-of-goods of these therapies. A plethora of microfluidic systems have also emerged in the last years for many cell culture applications. These systems provide increased process control due to their reduced dimensions and share many attributes of GMP-in-a-box systems: they are amenable for automation and monitoring, as well as single-use applications. Microfluidic cell culture technology may indeed provide new avenues for the automation and intensification of autologous CAR T manufacturing processes. In this thesis, a microfluidic device previously used for the perfusion culture and reprogramming of adherent stem cells was modified and adapted into a system for the expansion of CAR T cells in suspension, with integrated in-line oxygen monitoring. Microfluidic devices were designed, fabricated from cyclo olefin polymer and tested for the basic requirements of a CAR T cell expansion platform. This device, with a culture surface of 2 cm2 and an operating volume of 0.25 mL, incorporated two perfusion channels as well as two “side-channels” for seeding and harvesting. Robust device assembly and handling methods were also developed, with a special focus in reproducible operation. The final microfluidic device here described (Device C) allowed to achieve 10-fold CAR T cell expansion over four days, reaching 59x106 cell·mL-1 with 95% viability and comparable phenotype to GRex® controls. Although the total viable CAR T cell yield (8x106 cell), was insufficient for a single dose of CAR T treatment (which can require up to 108 cells), the high cell concentration achieved underpins the capacity of microfluidics for exquisite process control and potential for intensification of CAR T cell manufacturing processes. Automated, functionally closed, microfluidic processing units such as this may offer new sources of clinically viable CAR T cells.

Download activity - last month

Export as JSONCSVXML

Download activity - last 12 months

Export as JSONXMLCSV

Downloads by country - last 12 months

Export as XMLCSVJSON

Archive Staff Only

View Item