Document Type

Thesis

Degree Name

Master of Science (MSc)

Department

Biology

Program Name/Specialization

Integrative Biology

Faculty/School

Faculty of Science

First Advisor

Katelin W Spiteri

Advisor Role

Master's Thesis

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.

Convocation Year

2014

Convocation Season

Spring

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