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2004
LSU-HHMI Summer Undergraduate Research Program |
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 Jeffery
P. Reboul, Honor Ame Walesby, Veterinary Medicine
Transformation of Equine Myometrial Smooth Muscle Cells
in Culture
The pathophysiology of endotoxemia-induced preterm fetal expulsion
in mares is poorly understood. To further investigate this topic,
our research group has successfully cultured equine myometrial
smooth muscle cells (EMSMC) in monolayer. However, EMSMC in
monolayer exhibit prolonged doubling times, poor recovery from
cryogenic storage, and become senescent at passage 5 or 6. Transfected
cells have shorter doubling times, an infinite life span, and
allow research to be repeated over time as the same cells can
be maintained and their homogenous nature preserved. We hypothesized
that EMSMC can be transfected to provide a more reliable method
for the growth and maintenance of EMSMC for pharmacologic, physiologic,
and molecular investigation of endotoxemia-induced preterm fetal
expulsion in mares. Four finite cell lines were thawed, grown
to 70% confluence in monolayer, trypsinized, and placed into
six-well plates. Cells were transfected with pSV3 neo plasmid
containing SV40 early region and neomycin resistant genes. A
G418 selection antibody was used to select transfected cells.
At present all four of the cell lines are growing in the presence
of G418. The cell lines which form colonies will be considered
successfully transfected, grown to confluence, and cryogenically
preserved for future research projects.
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