Home › Pharmacology › Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays
Pharmacology JoVE (Open Access) Citable · DOI

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

DOI: 10.3791/55726-v
What you'll learn
  • ✓ Extract and dissociate mouse embryonic neurons for culture
  • ✓ Establish multi-electrode array recordings of neuronal network activity
  • ✓ Demonstrate stimulus-response training protocols in neuronal networks
  • ✓ Analyze time-dependent changes in spike frequency and network responses
Protocol

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

Difficulty
advanced
Total time
~3–4 days (dissection and plating ~2 hrs; culture maintenance 2–3 days before recording; recording session ~1 hr)
Model organism
Mouse (embryonic cortical neurons)
Biosafety
BSL-1

Steps

1
Remove embryos and extract brain tissue

Dissect embryonic mouse brain following IACUC-approved protocols. Extract intact brains for subsequent regional isolation.

▶ 01:04
2
Isolate frontal cortex from brain tissue

Remove frontal cortex regions under a dissection microscope. Collect cortical tissue for cell dissociation.

▶ 03:19
3
Dissociate cortical tissue into single cells

Apply enzymatic and mechanical dissociation to obtain a single-cell suspension. Prepare cell suspension for plating.

▶ 04:46
4
Plate neurons on multi-electrode arrays

Seed dissociated neurons onto pre-treated multi-electrode array devices. Maintain cultures in defined medium with regular media changes.

▶ 06:48
5
Perform visual inspection and record activity

Inspect cultures for cell viability and morphology. Connect to multi-electrode array system and record baseline and stimulus-evoked neuronal activity.

▶ 08:01
6
Analyze trained network response changes

Compare spike frequency and network response patterns between untrained and trained neuronal networks. Demonstrate significant alterations in electrophysiological properties.

▶ 09:24
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