Isolating Cranial Neural Folds From a Chick Embryo for a Neural Crest Cell Culture

Take a chick embryo supported within a filter paper frame and position it under a dissecting microscope.

Remove the vitelline membrane, a protective coating surrounding the embryo.

Identify the midbrain neural folds located posterior to the optic lobes and anterior to the cardiac crescent, the embryonic precursor of the mature heart.

The neural folds contain premigratory neural crest cells, which are multipotent stem cells that transform into migratory cells during development.

These migratory cells differentiate into neuronal and non-neuronal cells.

Excise the dorsal-most aspect of the neural folds and transfer the tissue to a culture dish containing buffer.

Next, place coverslips coated with an adhesion protein in culture plate wells containing media.

Transfer the neural folds onto the coated coverslips. Incubate to facilitate tissue attachment.

During culturing, migratory neural crest cells emerge from the neural folds onto the coated surface, establishing a neural crest cell culture.

Transfer an embryo to a clean dish containing Ringer's Pen-Strep solution. Gently swish the embryo back and forth to clear away any yolk that obscures the view, and exchange the Ringer's Pen-Strep