Co-culturing Glutamatergic Neurons and Pediatric High-Grade Glioma Cells

Begin with a polymer-coated microfluidic device comprising reservoir wells connected by channels and filled with culture medium.

Remove the medium from the reservoirs and seed human induced pluripotent stem cells.

The cells enter the channels and adhere to the coated surface.

Now, fill the reservoirs with a differentiation medium.

Growth factors in the medium induce stem cell differentiation into neuronal progenitor cells.

Next, add a medium containing specific nutrients and neurotrophic factors that induce neuronal progenitor maturation into glutamatergic neurons.

These neurons release the excitatory neurotransmitter glutamate from their presynaptic terminals, which bind to postsynaptic neuron receptors, generating excitatory signals.

Now, take pediatric high-grade glioma cells, cancer cells derived from aggressive brain tumors.

Add the cells to the reservoirs and incubate. The glioma cells enter the channels and adhere.

Gradually, these cells form functional synapses with the glutamatergic neurons, facilitating neuronal-glioma cell crosstalk and leading to neuronal hyperactivity.