Generating Neuromuscular Junctions Using a Microfluidic Device

Begin with a polymer-coated microfluidic device featuring interconnected wells carrying mirrored compartments that are connected via microgrooves.

Seed neural progenitor cells into the wells of one compartment.

Allow them to migrate into the interconnected channel and adhere to the polymer.

Overlay with a growth medium to promote neuronal maturation into motor neurons.

In the opposite compartment, seed the wells with muscle progenitor cells, which migrate and adhere to the interconnected channel.

Replace the medium with a differentiation medium, inducing the fusion of muscle progenitor cells and forming multinucleated muscle cells.

Add neuron-differentiation medium containing growth factors to the muscle cell-containing compartment.

Meanwhile, add a half volume of growth factor-free medium to the neural cell-containing compartment.

The growth factors and a higher volume of medium in the opposite compartment promote the motor neurons to extend their processes through the microgrooves.

These neuronal processes then connect with the muscle cells, forming neuromuscular junctions.