I am currently working to develop
plasmid vectors that will allow the Anseth Group to
study and manipulate gene expression in cells that are
used for creating tissues in the lab. The introduction
of exogenous genes into mammalian cells is a useful tool
for understanding the molecular mechanisms that underlie
different cellular phenotypes. For example, the
transfection of valvular interstitial cells (VICs) with
a reporter plasmid containing the alpha-smooth muscle
actin promoter that drives the expression of luciferase
allows researchers to quickly analyze the effects of
stimulatory growth factors on myofibroblast
differentiation. In addition, the transfection of VICs
with plasmids that over-express specific MAP Kinases
(both constitutively active and dominant negative forms)
can help to pinpoint the signaling pathways that are
involved in directing the myofibroblast phenotype. The
identification of pathways that direct differentiation
is vital to our goal of developing methods for
engineering tissues because our ability to create the
best environments for generating tissue is dependent on
our understanding of the mechanisms involved cellular
viability and differentiation.
In addition, I will be designing vectors that allow us to introduce exogenous
genes that actively induce stimulate cellular differentiation. By expressing
specific genes in human Mesenchymal Stromal Cells (hMSCs), for example, we hope
speed up the development of bone tissue in our 3-dimensional matrices. The
potential to use this technique for directing the differentiation of other
celltypes will be also pursued.
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