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The purpose of our research is to explore the effect of lead on neural crest cells. It is our belief that adding lead to post migratory NC cells will result in a higher rate of spontaneous neurite outgrowth than normal. We will explore this hypothesis by culturing NC cells in two different concentrations of lead-acetate and two control cultures of NC cells in serum.

Materials and Methods

Step 1:Neural Tube Removal

Eggs were kept in an incubator at 37oC for two days. Six two day old embryos were harvested from the eggs and the embryos were placed in petri dishes of Howard Ringer's solution. Using tungsten needles, we dissected out blocks of trunk from the embryos at the level of the last 10 somites, plus the unsegmented mesoderm anterior to Hensen's node.
The blocks were then transferred via pipet to a solution of 0.1% trypsin in distilled water (1:10 dilution). Modification of Dupin's protocol (1993) for removing the neural tube resulted in us leaving the blocks in the digestive solution for about five minutes. The blocks were then transferred to petri dishes with medium, where tungsten needles were used to pull off the excess mesoderm and somites surrounding the neural tubes, leaving only the neural tube.

©Cebra-Thomas, 2000

Last Modified: 5 May 2000

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