| Abstract |
Embryonic cell lines constitute an experimental tool for in vitro studies
on animal development as well as on more basic cell processes. These cells
retain all developmental potentials, allowing the modification of gene
expression by a precise control of the environmental conditions. Here we
report the initial characterization of two monoclonal embryonic cell lines
from Sparus aurata, SAEC-C1 and SAEC-F1. Cells were obtained from 6 hours
post-fertilized eggs and cultured in L-15 media, 10% FBS, 15.0 mM NaCl at
22 Celsius. After passing 20 times, a limit dilution was carried out and
single cell-derived cell lines were selected. The capacity of cell lines
to form embryoid bodies was evaluated by cultivation in non-adherent
plastic dishes as well as the ability to be transfected using EGFP and
DsRed plasmids. Moreover, gene expression was analyzed by RTqPCR. Cell
lines actively grow as adherent monolayers, displaying small and round or
polygonal morphologies. After cryopreservation and thawing, they show a
viability over 83%. Dissociated cells formed clusters and spherical
structures indicative of embryoid bodies. Both cell lines were
successfully transfected, being the fluorescence detected at 48-72 hours
after transfection. The expression of genes associated with stress (hsp70,
sod2, cat, gpx4), fatty acids metabolism, (fas, lpl, scd1a and delta6fad)
immune innate response (saBD, hep, lyz, ppar?, lox, cox2) and apoptosis
(Bcl2, Bax) was detected in both lines. Our results confirm that these
monoclonal embryonic cell lines could represent an important tool to
enlarge our knowledge on cellular processes as well as to perform in vitro
studies on genetic manipulation in sea bream.
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