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Differentiation of airway cholinergic neurons from human pluripotent stem cells for airway neurobiology studies

Goldsteen, P.; Sabogal Guaqueta, A.; Bos, S.; Kistemaker, L.; Van der Koog, L.; Eggens, M.; Halayko, A. J.; Dolga, A.; Gosens, R.

2022-03-26 neuroscience
10.1101/2022.03.23.485449 bioRxiv
Show abstract

Airway cholinergic nerves play a key role in airway physiology and disease. In asthma and other diseases of the respiratory tract, airway cholinergic neurons undergo plasticity and contribute to airway hyperresponsiveness and mucus secretion. We currently lack mechanistic understanding of airway cholinergic neuroplasticity due to the absence of human in vitro models. Here, we developed the first human in vitro model for airway cholinergic neurons using human pluripotent stem cell (hPSC) technology. hPSCs were differentiated towards mature and functional airway cholinergic neurons via a vagal precursor. Airway cholinergic neurons were characterized by ChAT and VAChT expression, and responded to chemical stimulation with changes in Ca2+ mobilization. Co-culture of hPSC-derived airway cholinergic neurons with airway smooth muscle cells enhanced phenotypic and functional characteristics of these neurons. The differentiation protocol we developed for human airway cholinergic neurons from hPSCs allows for studies into airway neurobiology and airway neuroplasticity in disease.

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