Isolating and Culturing Chick Embryonic Neurons on a Multi-Electrode Array
Transcription
Take a fertilized chicken egg containing an early-stage embryo.
Sterilize the shell and carefully open it.
Remove the embryo. Dissect and transfer the head to a culture plate containing a buffer. Remove the eyeball.
Make an incision and remove the skin layers, exposing the forebrain and midbrain.
Remove the inner membranous layer and blood vessels. Dissect and transfer the forebrain to a culture plate with a buffer.
Cut the tissue into smaller fragments and transfer them to a conical tube. Once the fragments settle, remove the buffer.
Add a proteolytic enzyme solution. Incubate to digest the tissue's extracellular matrix and loosen neurons.
Remove enzyme-rich media, wash with neurobasal media, removing any remaining enzymes.
Add neurobasal media. Mechanically dissociate the tissue to obtain a neuronal suspension.
Seed these neurons on a multi-electrode array system coated with an extracellular matrix. Add neurobasal media and incubate.
Neurons attach and extend projections, forming a neuronal network.
