Qi, John;
(2024)
Setting up a system to study the role of substrate stiffness on neural crest Epithelial to Mesenchymal Transition (EMT).
Masters thesis (M.Phil), UCL (University College London).
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Abstract
The neural crest is a highly migratory embryonic cell population known to differentiate into a multitude of various cell types such as the neurons, bone and pigment cells (Theveneau & Mayor, 2012), among many others. However, this diversity also requires them to migrate to various regions of the organism, leading to proper function. To do so, they must undergo a process known as Epithelial to Mesenchymal Transition (EMT). In the epithelial state, cell often adhere to neighbouring cells, maintaining an apical-basal cell polarity, and lacking cell motility. Yet, as they undergo EMT they become more migratory losing their cell-cell adhesion and apical-basal cell polarity. Although neural crest cell (NCC) EMT is widely studied and various genes and transcription factors have been identified, the process is still not completely understood. One factor that has recently been suggested to influence EMT is the mechanical properties of the substrate. A recent paper by Barriga et al. indicates that EMT can be inhibited by softening the underlying stiffness. To study the mechanism that is controlling EMT through stiffness, a robust and reproducible system needs to be developed. In this project, we aim to develop a system to study the role of how the substrate stiffness affects EMT of NCC in human cells in vitro. To do this, the precursors of NCCs, called neural border cells (NBC), are plated on polyacrylamide gels of various stiffnesses, and allowed to undergo EMT to form NCCs. Morphology, cell behaviour, gene expression and protein localisation were all analysed to determine if EMT has occurred. Observations of morphology and cell behaviour show cells on lower stiffness appearing more rounded and more epithelial than cells on a stiffer surface. The expression of EMT related genes such as Twist were shown to be upregulated while epithelial genes such as E-Cadherin were downregulated. Coupled with immunostaining also showed an upregulation of E-Cadherin expression, this provided enough validation to view this as a plausible model for studying EMT on different stiffnesses.
| Type: | Thesis (Masters) |
|---|---|
| Qualification: | M.Phil |
| Title: | Setting up a system to study the role of substrate stiffness on neural crest Epithelial to Mesenchymal Transition (EMT) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2024. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/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 UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10194596 |
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