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Research Project
Recent research has revealed that stem cells have the potential
to regenerate injured tissue, to serve as vehicles for gene
therapy, and to develop into organs for transplantation. They
have also been shown to be appropriate models for the study
of cellular differentiation and carcinogenesis. Stem cells are
defined by their ability to self-renew and differentiate into
various cell types. The mechanism of their differentiation is
not completely understood, but intercellular communication is
believed to be a critical component of the process. Toxicological
studies have indicated that certain environmental chemicals
can block cellular communication, implicating that stem cell
differentiation may also be affected. Here, we use an immortalized
stem-like cell line as a model to study the effects of environmental
toxicants on the cell-signaling pathways that control differentiation,
proliferation and apoptotic events.
The stem-like cell line, SVG, used in this study is derived
from a human fetal brain and has been characterized as astroglial
in origin. These cells have been shown to differentiate and
project neurite-like structures in the presence of cAMP inducers.
One of our aims is to construct a custom microarray
containing genes relevant to the differentiation of SVG cells.
A custom array would allow us to screen genes of interest on
a uniform platform and minimize data analysis. To date, about
3000 genes that are expressed in the SVG cells have been identified
by screening several commercially available arrays. In addition
to the genes found experimentally, potential genes of interest
were also chosen based on published literature. Expressed sequence
tags representing these genes were obtained from the IMAGE consortium.
Following PCR amplification and purification, the DNA representing
each gene was printed onto glass slides.
My current project involves using the custom
arrays to establish an expression profile for these cells when
induced to differentiate in the absence of environment toxicants.
Once the baseline expression levels have been determined, these
cells will be stimulated to differentiate in the presence of
environmentally relevant chemicals and gene expression will
be monitored. In addition to expression profiling, I am also
responsible for the construction of a second version of the
SVG microarray, which will contain DNA sequences representing
a larger set of genes. We anticipate that this study will further
our understanding of stem cell differentiation and the effects
of environmental toxicants on this process.
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