Helenes Gonzalez, Citlali;
(2019)
Novel tools and techniques for studying human neural cell behaviour and development.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
During neural development, human neural stem cells (hNSCs) self-renew and generate the neurons and glia of the central nervous system (CNS). The brain extracellular matrix (bECM) provides biomechanical and chemical cues, which regulate hNSCs behaviour. Many fundamental questions about hNSCs and CNS development still remain unanswered. Emerging technologies can provide invaluable insights for understanding hNSCs environment and establishing reproducible neural systems. The aim of the project was to investigate new approaches to model human neural tissue in vitro that will facilitate studying CNS behaviour and development, with a particular focus on how human neural cells respond to different environments. Specific objectives included: 1) investigating hNSCs following bio-electrospray (BES) to assess BES potential for developing neuro-scaffolds; 2) evaluating the effects of nanodiamonds (NDs) with different surface modifications on hNSC growth and differentiation; 3) characterising porcine brain decellularisation protocols and the human developing bECM for future development of bECM-based scaffolds. As shown by cellular and molecular analysis, BES did not affect hNSC viability and differentiation, but highlighted the need for suitable sprayable scaffolds. NDs sustained neuronal differentiation and neurite outgrowth, regardless of surface chemistry, as indicated by neurite tracing and RT-PCR, supporting their suitability for certain biomedical applications. Conditions were established to advance standardisation of decellularisation and solubilisation of porcine bECM. Additionally, a protein quantification protocol was developed. Porcine bECM was found to potentiate hNSCs proliferation in presence of laminin, but not alone. Human foetal bECM was then investigated by mass spectrometry-based proteomics, and 73 matrix proteins were identified. The brain matrisome composition appeared fairly stable during development, with the main changes observed between 10 and 20 weeks of gestation. This multidisciplinary study provides novel insights for developing controlled human CNS models in vitro that are much needed for biomedical research and basic understanding of human brain function and development.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Novel tools and techniques for studying human neural cell behaviour and development |
| 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 |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10088430 |
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