1 research outputs found
Establishment, Characterization, and Toxicological Application of Loggerhead Sea Turtle (Caretta caretta) Primary Skin Fibroblast Cell Cultures
Pollution is a well-known threat
to sea turtles but its impact
is poorly understood. In vitro toxicity testing presents a promising
avenue to assess and monitor the effects of environmental pollutants
in these animals within the legal constraints of their endangered
status. Reptilian cell cultures are rare and, in sea turtles, largely
derived from animals affected by tumors. Here we describe the full
characterization of primary skin fibroblast cell cultures derived
from biopsies of multiple healthy loggerhead sea turtles (Caretta caretta), and the subsequent optimization
of traditional in vitro toxicity assays to reptilian cells. Characterization
included validating fibroblast cells by morphology and immunocytochemistry,
and optimizing culture conditions by use of growth curve assays with
a fractional factorial experimental design. Two cell viability assays,
MTT and lactate dehydrogenase (LDH), and an assay measuring cytochrome
P4501A (CYP1A) expression by quantitative PCR were optimized in the
characterized cells. MTT and LDH assays confirmed cytotoxicity of
perfluorooctanoic acid at 500 μM following 72 and 96 h exposures
while CYP1A5 induction was detected after 72 h exposure to 0.1–10
μM benzo[<i>a</i>]pyrene. This research demonstrates
the validity of in vitro toxicity testing in sea turtles and highlights
the need to optimize mammalian assays to reptilian cells