Raimes, William N.M.;
(2019)
An Automated Microfluidic Perfusion System for the Derivation of Induced Pluripotent Stem Cells.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
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.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | An Automated Microfluidic Perfusion System for the Derivation of Induced Pluripotent Stem Cells |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2019. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10071157 |
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