Analyzing Tooth Germ and Trigeminal Ganglia Interactions Using a Microfluidic Co-culture System

Take a microfluidic device coated with a synthetic polymer and an adhesion protein.

The device consists of wells and interconnected culture chambers separated by microgrooves.  

The chambers contain punched holes for tissue placement.

Obtain the trigeminal ganglia, a collection of trigeminal neuron cell bodies, and tooth germs, prenatal structures that develop into adult teeth, from a mouse embryo.

Place the tissues in the holes and add the respective culture media.

Nutrients and vitamins in the tooth germ media facilitate tooth germ survival during culturing.

The coated device surface and growth factors in the trigeminal ganglia media facilitate neurite and axonal outgrowth.

The axons reach the tooth germ chamber through the microgrooves.       

The tooth-derived signals lead to neuronal repulsion, allowing the axons to surround the tooth germ without innervating it.

Wash the culture chambers with buffer and fix the tissues with a fixative.

The co-culture is now ready for further analysis.