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Title: An automated microfluidic perfusion system for the derivation of induced pluripotent stem cells
Author: Raimes, William N. M.
ISNI:       0000 0004 7964 9266
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2019
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The advent of induced pluripotent stem cells (iPSCs) i s a paradigm shift for the field of personalised medicine. iPSCs have immense potential as an ethical and non - immunogenic source for autologous transplant or as disease models for drug screening and tox icology. A key bottleneck for iPSC clinical translation is the expensive and lengthy manufacturing process from reprogramming to pre - transplant screening. Automated bioprocessing systems have emerged in recent years to standardise and reduce costs of cell therapy . One promising avenue involves the use of microfluidic culture systems for reduce d resource use and precision control. In this thesis, a microfluidic device previously designed for uniform perfusion culture of adherent stem cells wa s developed into a plat form for reprogramming on - chip that integrates transfection, perfusion culture and online monitoring. On - chip chemical transfection was achieved using a switching valve to automate small volume injection of episomal vectors. A closed - channel perfusi on chip fabricated from cyclic olefin polymer was designed in parallel with a novel clamping frame to simplify assembly and improve robustness. A suitable reprogramming protocol compatible with Good Manufacturing Practice was verified at bench scale , before being downscaled in to the perfusion chip for a proof - of - concept automated reprogramming of human dermal fibroblasts. Comparison of microscale reprogramming in static and perfused conditions revealed no significant difference in efficiency of colony formation at a chamber dilution rate of 0.144 volumes .min - 1 (chamber volume = 10.5 μ l). iPSCs manually isolated from this system and expanded in well plates were characterised as pluripotent cells, able to differentiate into three germ layers and displayin g normal karyotype. An automated microfluidic processing unit such as this is the first step toward a cost - efficient , commercially relevant source for clinically viable iPSC s . It is also a useful tool for the development of process analytical technologies for online visual monitoring to reduce requirement for downstream quality control.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available