Recording Electrophysiological Activity of the Neuronal Network in a Neuron-Astrocyte Co-culture

Begin with a multi-electrode array, or MEA, plate. Each well contains a centrally positioned array of electrodes.

The array is coated with a culture substrate to facilitate cell adhesion.

Add a suspension of motor neurons and astrocytes as a droplet over the array.

Incubate to allow cell attachment, then add a warm co-culture medium supplemented with a small molecule that maintains cell viability.

During incubation, astrocytes secrete factors that promote neuronal maturation and synapse formation, resulting in a neuronal network.

Place the MEA in a recording device under physiological conditions to monitor neuronal activity.

Neurons transmit electrical signals, called action potentials, that travel along their axons.

Upon reaching the synapse, the signal triggers neurotransmitter release, propagating the signal to the next neuron.

Electrodes within each well extracellularly record the signals generated by synaptically connected neurons.

The detection of synchronized rhythmic electrical signals confirms the establishment of a neuronal network.

On the day of plating or before, start coating the 24-well MEA plates by first diluting PLO in water or PBS. Then, add 15 to 20 microliters o