This study demonstrates a method for recording spontaneous electrophysiological activity from a neuron-astrocyte co-culture using multi-well micro-electrode arrays (MEAs). Human induced pluripotent stem cell-derived neurons are co-cultured with rat astrocytes to support network maintenance.
Begin with a neuron-astrocyte co-culture in a multi-well micro-electrode array or MEA.
Each well contains a network of neurons derived from human induced pluripotent stem cells, or hiPSCs. The rat astrocytes provide support for maintaining the network.
The base of the wells contains a grid of tiny electrodes.
Place the MEA in a recording device to capture the spontaneous electrophysiological activity of the neuronal network.
Flow a humidified gas mixture into the culture to prevent media evaporation and stabilize the pH, ensuring cell viability.
Neurons transmit signals via electrical impulses, known as action potentials, that travel along their axons.
Upon reaching the synapse, the impulse triggers the release of neurotransmitters, which propagate the signal to the next neuron.
The electrodes within each well simultaneously record action potentials generated by multiple synaptically connected neurons.
The spontaneous electrophysiological activity of the neuronal network confirms the differentiation of hiPSCs into neurons.
Acquire the data at 1,200 times amplification and sample the signal at 10 kilohertz using the MCS data acquisition card. Record 20 minutes of electrophysiological activity of hiPSC-derived neurons cultured on MEAs. During the recording, maintain the temperature at 37 degrees Celcius and prevent medium evaporation and pH changes by delivering a constant slow flow of humidified gas onto the MEAs. After that, analyze the data using a custom software package.
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