Differentiation of Neural Stem Cells to Neurons Using a Compartmentalized Microfluidic Device

Take a microfluidic device consisting of reservoir wells connected to two compartments, a somatic and an axonal, separated by a microgroove barrier.

The compartments and microgrooves are coated with a cell culture substrate to facilitate cell adhesion.

Add a neural stem cell medium to each well and incubate to precondition the device for cell culture.

Discard the media. Add neural stem cells, or NSCs, to the wells of the somatic compartment and allow the cells to adhere.

Add the NSC medium to each well and incubate in a humidified chamber to prevent media evaporation. During incubation, the NSCs proliferate.

Replace the NSC medium with a neural differentiation medium and incubate.

The small molecules in the medium induce the differentiation of NSCs into neurons within the somatic compartment, with their axons extending through the microgrooves into the axonal compartment.

The differentiated neurons with extended axons are ready for further assays.

To begin, dissolve poly-l-ornithine in cell culture-grade distilled water to make 600 microliters of solution per chip. Aspirate the remaining PBS from the wells, making sure that the pipette tip is away from the channel opening. Load 150