• medil ganglionic eminence progenitors
• Alzheimer's disease
• basal forebrain cholinergic neurons
• Parkinson's disease with dementia
• dementia with Lewy bodies

Alzheimer’s disease (AD), dementia with Lewy bodies (DLB) and Parkinson’s disease with dementia (PDD) are common neurodegenerative diseases, mainly caused by the presence of intracellular neurotoxic aggregates (Lewy bodies or Aβ-oligomers), affecting, among other brain regions, the basal forebrain cholinergic system. Accumulation of toxic protein aggregates in the Nucleus Basalis Magnocellularis (NBM) is usually accompanied by degeneration of acetylcholine-releasing neurons and cholinergic denervation, leading to the typical disabling symptoms of dementias such as visual hallucinations, memory loss and attention fluctuation. Current available treatments, such as cholinesterase inhibitors, serotonin agonists, and deep brain stimulation (DBS) provide temporary symptomatic relief without counteracting the cell loss. Furthermore, these treatments require the presence in situ of residual functional neurons and therefore are not effective when the cells are completely degenerated. In this context, cell replacement therapy is a promising approach to repair the damaged cholinergic system and restore the normal brain circuitry, thereby alleviating the cognitive deficits.
The aim of this thesis work was to optimize in vitro patterning of human pluripotent stem cells (hPSCs) towards a basal forebrain cholinergic neurons (BFCN) progenitor fate, to generate a homogeneous product which may be used as a cell therapy for treatment of dementias with cholinergic degeneration. To this aim, I explored combinations of different morphogens to generate authentic BFCNs while suppressing differentiation of non-BFCN neuronal populations with similar developmental origin, such as the GABAergic interneurons. In particular, I investigated the effect of two maturation factors, BMP9 and NGF, that are known to play a central role in the specification of BFCNs, investigating long-term maturation of our cultures in order to assess their potential for in vitro disease modelling.