Generation of disease-relevant neurons from human pluripotent stem cells
Author(s)
Arber, Charles
Type
Thesis
Abstract
This thesis describes investigations into exogenous factors that influence fate-choices during
human pluripotent stem cell (PSC) differentiation in vitro. I describe novel growth factor
environments and small molecule regimes that provide patterning signals enabling directed
differentiation of human PSCs towards biomedically relevant neurons. I provide evidence that the
TGFβ growth factor Activin can promote ventral telencephalic differentiation. Small adjustments in
Activin administration can produce an over-representation of forebrain derived medium spiny
neurons and cortical interneurons, with significance in Huntington’s disease and epilepsy
respectively. Additionally, I manipulate the earliest growth factor environments of PSC
differentiation to lead to broad changes in rostro-caudal patterning. Inhibiting FGF signalling leads to
a midbrain-like phenotype that can be further differentiated towards a ventral midbrain
dopaminergic fate, a cell type that degenerates in Parkinson’s disease. Our novel manipulations and
differentiation protocols provide insights into early events in human development, which would
otherwise be impossible to study. This logic can also be applied to investigate diseased states during
human development and ageing via use of disease-specific cell lines. The mature neurons produced
may provide a tool that can be applied to large-scale drug screening assays and toxicology testing as
well as having the potential for cell based therapies in future years.
human pluripotent stem cell (PSC) differentiation in vitro. I describe novel growth factor
environments and small molecule regimes that provide patterning signals enabling directed
differentiation of human PSCs towards biomedically relevant neurons. I provide evidence that the
TGFβ growth factor Activin can promote ventral telencephalic differentiation. Small adjustments in
Activin administration can produce an over-representation of forebrain derived medium spiny
neurons and cortical interneurons, with significance in Huntington’s disease and epilepsy
respectively. Additionally, I manipulate the earliest growth factor environments of PSC
differentiation to lead to broad changes in rostro-caudal patterning. Inhibiting FGF signalling leads to
a midbrain-like phenotype that can be further differentiated towards a ventral midbrain
dopaminergic fate, a cell type that degenerates in Parkinson’s disease. Our novel manipulations and
differentiation protocols provide insights into early events in human development, which would
otherwise be impossible to study. This logic can also be applied to investigate diseased states during
human development and ageing via use of disease-specific cell lines. The mature neurons produced
may provide a tool that can be applied to large-scale drug screening assays and toxicology testing as
well as having the potential for cell based therapies in future years.
Date Issued
2012
Date Awarded
2013-02
Copyright Statement
Attribution NoDerivatives 4.0 International Licence (CC BY-ND)
Advisor
Li, Meng
Sponsor
Medical Research Council (Great Britain) ; EuroStemCell ; Imperial College London
Publisher Department
Institute of Clinical Science
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)