A NOVEL STEM-CELL BASED CO-CULTURE DEVICE TO STUDY NEUROMUSCULAR DISORDERS

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disease characterised by the degeneration of neuromuscular junctions and motor neurons. To gain insight into mechanisms of ALS and other neuromuscular disorders, it is crucial to develop a relevant in vitro system to model nerve-muscle connectivity. We established a compartmentalised model to co-culture motor neurons, astrocytes and muscle. Individual neural cell types were derived from mouse embryonic stem cells and purified while muscle was derived from chick embryonic primary myoblasts. The cells were assembled in microfabricated devices that segregate the cell bodies of neural cells from muscle fibres to mimic the normal anatomy of neuromuscular connectivity. Motor neurons were equipped with Channelrhodopsin-2, enabling their selective optogenetic activation. Using this device, we have achieved long-term co-cultures of muscle and neural tissues. We have also demonstrated the formation of neuromuscular junctions and their functionality.