Microfluidic device for three dimensional and compartmentalized coculture of neuronal and muscle cells, with functional force readout

A method comprising:

• seeding a second culture compartment with muscle cells to enable growth of a muscle bundle, the muscle bundle wrapping around compliant pillars, the second culture compartment forming part of a microfluidic device;
• and
• seeding a first culture compartment with neuronal cells to enable growth of an axon to extend from the first culture compartment through a buffer compartment to the second culture compartment and form a three-dimensional neuromuscular junction with the muscle bundle;
• wherein the microfluidic device is for coculture of the muscle cell and the neuronal cells for innervating the muscle bundle, the microfluidic device comprising:
• a coculture chamber or a plurality of coculture chambers, each of said chamber comprising:
• the first culture compartment including one or more retaining features, the first culture compartment having a width of at least 100 μm;
• the second culture compartment including the one or more compliant pillars, the second culture compartment having a width of at least 100 μm;
• and
• the buffer compartment separating the first compartment and the second compartment by a distance of at least about 200 μm;
• wherein the compliant pillars are deflectable to measure force generated by the muscle bundle and the one or more compliant pillars have a height of at least 50 μm.