Cellular and molecular characterisation of human pluripotent stem cell derived medium striatal neurons

Prapaiwongs, Parinda 2025. Cellular and molecular characterisation of human pluripotent stem cell derived medium striatal neurons. PhD Thesis, Cardiff University. Item availability restricted.

Preview	PDF - Accepted Post-Print Version Download (279MB) | Preview
	PDF (Cardiff University Electronic Publication Form) - Supplemental Material Restricted to Repository staff only Download (420kB)

Abstract

GABAergic medium spiny projection neurons (MSNs) comprise the majority of cell populations in the striatum along with different types of interneurons (INTs). These projection neurons are profoundly degenerated in Huntington's disease (HD) and aberrant striatal development has been associated with neurodevelopmental and neuropsychiatric disorders. Although Activin A-based differentiation protocol has enabled striatal neuron derivation from human pluripotent stem cells (hPSCs), the molecular authenticity and contribution of INTs to MSN development remain insufficiently characterised. Current validation of hPSC-derived striatal cultures has relied on limited set of canonical markers assessed at bulk and protein levels. This approach offers only partial insight into cellular heterogeneity, developmental trajectories, and fidelity to native human striatal development. While INTs constitute a substantial proportion of the human striatum compared to rodent striatum, their influence on the development of MSN has not been systematically examined in human in vitro models. This thesis characterised hPSC-derived striatal cultures by using single-cell RNA sequencing (scRNA-seq), integrating comparative bioinformatics with human fetal lateral ganglionic eminence (LGE) datasets, and employing three-dimensional organoid and assembloid systems combined with immunocytochemistry and singlenucleus RNA sequencing (snRNA-seq). The findings demonstrated that Activin A-patterned hPSC-derived striatal cells recapitulate key molecular and developmental features of human fetal LGE, including progressive specification of direct and indirect pathway MSNs. However, they exhibited altered developmental timing and asynchronous maturation compared to the human fetal LGE. Potential novel stage- and subtype-associated molecular markers and coordinated regulatory programs governing MSN differentiation were identified. Furthermore, 3D co-culture studies indicated that INTs exert non-cell-autonomous influences on MSN maturation dynamics without altering terminal fate. ii Collectively, this thesis established a robust molecular reference for hPSC-derived striatal cultures, advances understanding of human MSN and INT development, and provides a foundation for modelling basal ganglia disorders and informing future regenerative strategies for HD.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Schools > Biosciences
Subjects:	Q Science > Q Science (General)
Date of First Compliant Deposit:	12 May 2026
Last Modified:	12 May 2026 14:27
URI:	https://orca.cardiff.ac.uk/id/eprint/186946

Actions (repository staff only)

Edit Item