Objective
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.
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