This project proposes a novel and simple technique to pattern glia and neurons. This study investigates various microfabrication friendly patterning systems and develops a highly consistent, straightforward and cost effective cell patterning scheme based on two common ingredients: the polymer parylene and horse serum. The first part of this work describes the new patterning method and highlights its capabilities. Stripes of parylene on silicon thermal oxide (SiO<sub>2</sub>) were fabricated in the cleanroom, cleaned and subsequently activated by a simple deep and rinse process in horse serum. Glia and neurons from dissociated rat hippocampi were cultured on the patterns and imaged after 7 DIV. Standard cell culture and immunofluorescent imaging protocols were used. A comparison between experimental and control samples indicate that the protein load in horse return serum is able to alter parylene into a cytophylic substrate. The second part of this project focuses on the optimisation of the parylene based patterning method. Aspects, such as the sustainability of the cell patterns beyond the first week and the effect of glia division on the quality of the patterns are investigated. Furthermore, the reduction of the immersion time of the substrates in the horse serum is explored while a behavioural model for the cells during their first week in culture is proposed. Submersion in an alternative serum is examined while the effects of UV radiation on the parylene are revealed not to be contributing to cell patterning. In the last part of this study an attempt is made to analyse the parylene and thermal oxide surfaces before and after immersion in the horse serum. An initial theory explaining the preference of cells towards parylene is disproved, while three probable hypotheses are proposed. Evidence from XPS analysis and electrophoresis gels suggests that the amount and conformation of proteins on the parylene and thermal oxide substrates might be responsible for inducing glial and neuronal patterning.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:649469 |
| Date | January 2008 |
| Creators | Delivopoulos, Evangelos |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/13611 |
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