From high-content to super-resolution investigation of cell behaviour on nanostructured surfaces

Reynolds, Paul M (2015) From high-content to super-resolution investigation of cell behaviour on nanostructured surfaces. PhD thesis, University of Glasgow.

Due to Embargo and/or Third Party Copyright restrictions, this thesis is not available in this service.

Abstract

The environment in which cells find themselves is a complex, three dimensional one which provides a variety of inputs and cues capable of controlling and guiding cell behaviour. These environmental signals take a fertilised egg through development to become an adult human being made up of trillions of cells. As such, the power of environmental cues to provide context and guidance to cell behaviour cannot be understated. Without attempting to directly mimic the in vivo environment, it has been shown that micro- and nanostructured surfaces can influence cell behaviour when we try and engineer biology in vitro. Identification and optimisation of powerful topographies is, however, tedious, and so this thesis provides techniques to expedite the discovery of new and potent surfaces to drive cell behaviour. A new fabrication technique has been developed which allows for the fabrication of gradients of feature height at both the micro- and nanoscale. This involves the use of plasma polymer gradients as novel etch masks alongside existing lithographic techniques. After fabrication and mass replication by injection moulding, use of these surfaces as platforms for the high-content screening of cell response is demonstrated. These can be considered high-content due to both the range of surface structures on a single sample, and also the microscopy techniques used to investigate cell response. Distinct cell types were found to respond differently to topographical cues, exhibiting varying degrees of alignment, proliferation, and organisation in both mono- and co-culture systems. A new cell culture device has also been developed and patented which ensures that screening experiments begin with an accurate and repeatable distribution of cells across the high content array. The impact of uneven cell seeding on studies involving stem cell differentiation was also investigated – showing the importance of improved control. Finally, the interaction of cells with such nanostructured surfaces is investigated using new super-resolution microscopy techniques. New methods are presented for the correlation of multiple nanoscale imaging techniques to view cell-nanostructure interactions with unprecedented resolution. This reveals insights into the way in which the cellular substructure is being modulated by underlying nanotopography. Indeed, it paints a picture which is remarkably different to the structure observed under a standard widefield microscope over the past 10 years.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Keywords: High Throughput, cell engineering, gradient, nanofabrication
Subjects: Q Science > Q Science (General)
Colleges/Schools: College of Science and Engineering > School of Engineering > Biomedical Engineering
Funder's Name: UNSPECIFIED
Supervisor's Name: Gadegaard, Prof. Nikolaj and Riehle, Dr. Mathis
Date of Award: 2015
Embargo Date: 15 June 2018
Depositing User: Mr Paul Reynolds
Unique ID: glathesis:2015-6490
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 30 Jun 2015 12:19
Last Modified: 09 Jul 2015 14:39
URI: http://theses.gla.ac.uk/id/eprint/6490

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