Abstract
Understanding neurodegenerative conditions such as amyotrophic lateral sclerosis (ALS) requires a supply of cells for in vitro modelling and drug screening. Motor neurons derived from induced pluripotent stem cells (iPSC) provide an unlimited source of such cells and can potentially facilitate precision medicine approaches to compound testing when derived from somatic cells from patients. Furthermore, in co-culture with iPSC-derived muscle cells, a fully human preclinical screening model of the neuromuscular Junction is possible. However, iPSC-derived motor neurons traditionally required up to six weeks maturation to exhibit relevant functional phenotypes.
We demonstrate use of an in vivo environment-mimicking supplement, to produce a more physiological, modified growth medium for maturation, which reduces the time to reach functional maturity from six weeks to 10 days. Cells showed typical morphology with abundant neurites and were positive for expression of the mature motor neuron marker SMI-32 along with HB9, ChAT and Isl-1, as assessed by immunocytochemistry.
Recordings using a 48-well multi-electrode array (MEA) demonstrate synchronised spontaneous firing at a similar timepoint. Motor neurons matured using the accelerated protocol formed junctions with muscle cells in a microfluidic co-culture system, as demonstrated by staining with fluorescently labelled alpha-bungarotoxin. The phenotype of motor neurons from an ALS patient was also investigated.
These rapidly matured cells provide an invaluable tool for research into neuromuscular disease and a model allowing a higher throughput for compound screening.
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