Assessing Glutamatergic Neurons and Glioma Cells Interactions in a Co-Culture

Begin with a laminin-coated, compartmentalized microfluidic device positioned over a patterned multielectrode array or MEA.

The device carries cultured iPSC-derived cortical glutamatergic neurons adhered to microelectrodes of an MEA.

These neurons communicate via electrical signals. Record these signals as baseline neuronal activity.

Next, seed the device with glutamate -rich pediatric high-grade glioma cells, a type of brain tumor cell, and incubate.

Over time, glioma cells enter the channel and attach, establishing the co-culture. Later, glioma cells release glutamate, an excitatory neurotransmitter.

The released glutamate interacts with neuronal receptors, leading to an influx of calcium and sodium ions that causes neuronal membrane depolarization.

This induces neuronal excitability and enhances neuronal activity.

Re-record the electrical signals, and a significant increase after co-culture indicates successful neuron and glioma cell interaction.

Seed the trypsinized UW479 and BT35 cells on top of the mature adherent glutamatergic neurons in each dedicated microfluidic device. Maintain the co-cultures for two days under a controlled environment at 37 degrees Celsius and 5% carbon dioxide with glutamatergic neuron D11 and onward medium.

Count pediatric high-grade glioma cells using the microscopic pictures analyzed with the image analysis software to assess their viability, and calculate the percentage of cells. Place the MFD in the recording device, and use a commercially available software to perform the electrophysiological recording of glutamatergic neurons on the 21st day.

Before seeding pediatric high-grade glioma cells, perform a second electrophysiological recording of the differentiated glutamatergic neurons cultured as control, parallel to the co-culture.