| Abstract |
This thesis describes the establishment, characterization and potential
toxicological applications of YPF5, a yellow perch cell line. YPF5 is a
fibroblastic cell line derived from the caudal fin of yellow perch. This
cell line has been maintained for over 2 years. Cultures have been
passaged over 60 times, and successfully cryopreserved and thawed. YPF5 is
grown at room temperature in Leibovitz-15 (L-15) media, supplemented with
10% fetal bovine serum and 3% penicillin-streptomycin. The origin of YPF5
has been confirmed as Perca flavescens through 'DNA Barcoding' and
reaffirmed through Karyotyping (2n = 48) as per literature reported
karyology for percidae including the yellow perch. YPF5 tested positive
for alkaline phosphatase and negative for beta-galactosidase, markers of
stem cell-like characteristics and senescence, respectively, providing
evidence for a continuous cell line. Immunofluorescence staining with
mesodermal cell markers (vimentin and collagen type 1) also confirmed the
fibroblastic origin of the cells. In agreement with related literature,
the morphological response of YPF5 exposed to cortisol made cells more
epithelial-like, and exposure to ascorbic acid induced extracellular
collagen secretion. YPF5 responded to different contaminants (naphthenic
acid, copper sulphate, ammonium chloride and OSPW samples) in a dose
dependent manner. Evaluation of possible remediation methods using
bentonite and UV radiation to ameliorate toxicity of oil sands chemicals
showed promise in vitro with YPF5 cells and could be investigated further.
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