Gogolou, Antigoni ORCID: 0000-0002-3581-2426 (2022) Dissecting Neural Crest Ontogenesis and Axial Specification in Vitro Using Human Pluripotent Stem Cells. PhD thesis, University of Sheffield.
Abstract
Neural crest (NC) is a multipotent embryonic population integral to vertebrate development owing to its broad contribution to an array of derivatives including peripheral ganglia and the craniofacial skeleton. The NC cells originate from the dorsal tip of the neural tube along the anteroposterior axis and based on their positional identity, which is coupled with their ability to generate distinct cell types, are subdivided into cranial, cardiac and vagal, trunk and sacral NC in an order that corresponds to the adjacent regions of the spinal cord. The importance of NC integrity is demonstrated by the wide spectrum of NC-related pathologies including tumours, termed neurocristopathies, that predominantly arise from defects in the development of NC and their derivatives in multiple tissues. Our knowledge of NC ontogenesis and pathophysiology primarily comes from studies with animal models, however, the model organisms don’t always recapitulate human development. Thus, human pluripotent stem cells (hPSCs) are an attractive platform to study NC ontogenesis and pathogenesis in human context in vitro. Several studies using directed differentiation have decoded the signalling requirements for cranial NC specification from hPSCs, but the signals and transcription factors that drive the acquisition of a posterior axial identity that corresponds to vagal and trunk levels in in vitro-derived NC have been less studied. Recently, it was demonstrated that vagal NC cells are derived from cranial crest through the action of the caudalising factor retinoic acid (RA), whereas their trunk counterparts are generated downstream of a neuromesodermal progenitor (NMP) population. The work presented here defines Wnt along with cell density as critical factors for vagal NC specification in vitro and demonstrates the ability of RA-induced vagal NC to give rise to their enteric nervous system (ENS) derivatives upon further differentiation using two different hPSC lines. It also shows that Notch pathway inhibition as part of the culture regimen enhances neural and glial differentiation in ENS cultures. Additionally, it reveals a previously unknown role of the pro-mesodermal factor TBXT together with Wnt signalling effectors in regulating, through chromatin remodelling, the adoption of posterior axial identity in NMP-derived trunk NC. We believe that these insights contribute to our knowledge of posterior patterning in post-otic NC and they can be applied in the future to optimise NC differentiation strategies with practical implications in biomedicine.
Metadata
Supervisors: | Tsakiridis, Anestis and Andrews, Peter |
---|---|
Related URLs: |
|
Keywords: | hPSCs; Neural crest; anteroposterior patterning; enteric nervous system; neuromesodermal progenitors; TBXT |
Awarding institution: | University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) The University of Sheffield > Faculty of Science (Sheffield) |
Depositing User: | Ms Antigoni Gogolou |
Date Deposited: | 30 May 2023 08:28 |
Last Modified: | 30 May 2023 08:28 |
Downloads
Final eThesis - complete (pdf)
Embargoed until: 30 May 2024
Please use the button below to request a copy.
Filename: Antigoni Gogolou_thesis after corrections_v1_final.pdf
Supplementary Material
Embargoed until: 30 May 2024
Please use the button below to request a copy.
Filename: Combined Stacks_phase_eGFP.avi
Description: Time lapse video 1 of ENS progenitors_Phase contrast_eGFP
Supplementary Material
Embargoed until: 30 May 2024
Please use the button below to request a copy.
Filename: Combined Stacks3_phase_egfp.avi
Description: Time lapse video 3 of ENS progenitors_Phase contrast_eGFP
Supplementary Material
Embargoed until: 30 May 2024
Please use the button below to request a copy.
Filename: Combined Stacks 2_Phase_eGFP.avi
Description: Time lapse video 2 of ENS progenitors_Phase contrast_eGFP
Export
Statistics
Please use the 'Request a copy' link(s) in the 'Downloads' section above to request this thesis. This will be sent directly to someone who may authorise access.
You can contact us about this thesis. If you need to make a general enquiry, please see the Contact us page.