One of the first methods for the derivation of BFCNs from hESCs was based on the stimulation using diffusible ligands present in the MGE at developmentally relevant time periods .
Additional expression of NK2 homeobox 1 (NKX2-1) and ISL LIM homeobox 1 (ISL1) is associated with the development of forebrain cholinergic neurons and coexpression of p75 receptor demonstrated the presence of BFCNs. Furthermore, these differentiated neuronal cells produce [alpha]3, [alpha]4, and [alpha]7 subunits of nicotinic acetylcholine receptor and muscarinic acetylcholine receptor subtypes, M1, M2, and M3, that are displaying an important role in hESCs-derived cholinergic neurons .
Among the general cortical impairment, BFCNs are one most affected cell type in AD.
Furthermore, it can be used for the development of cell replacement therapy for Alzheimer's disease and high-throughput screening for agents that promote BFCNs survival.