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Volume 8
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X
Biotech Congress 2018 & Enzymology 2018
March 05-07, 2018
JOINT EVENT
20
th
Global Congress on
Biotechnology
3
rd
International Conference on
Enzymology and Molecular Biology
&
March 05-07, 2018 London, UK
Study of the potential use of antifreeze proteins of
Deschampsia antarctica
in the cryopreservation of
Salmo salar
spermatozoa
Short S, Farías J G, Díaz R
and
Bravo L A
Universidad de La Frontera, Chile
C
ryopreservation allows to preserve genetic resources in aquatic species, such as Atlantic salmon (
Salmo salar
). However,
freezing may cause cell damage affecting the sperm quality. New procedures including antifreeze proteins (AFPs) seem
to improve sperm quality after cryopreservation. AFPs have the ability to bind to ice crystals inhibiting their growth, and ice
recrystallization (IRI)
in vitro
.
Deschampsia antarctica
is a freezing tolerante vascular plant species (LT50 -27°C) exhibiting
apoplastic antifreeze activity. We hypothesize that AFPs from
D. antarctica
favor the sperm quality of cryopreserved
S.
salar
spermatozoa. The aim of this work is to evaluate cryoprotection of AFPs from
D. antarctica
in
S. salar
spermatozoa.
Cryopreservation of S. salar spermatozoa has been made with a standard freezing medium (C
+
) and different treatments with
protein extracts (20 μg/ml) of
D. antarctica
supplemented with permeating, DMSO 1.3 M, glucose 0.3 M, and non-permeating,
BSA 2% w/v cryoprotectants. Post-thawing plasma membrane integrity (PMI) by SYBR-14/PI and mitochondrial membrane
potential (MMP) by JC-1 markers were assessed using flow cytometry. Thawed cells in the presence of protein extracts from
D. antarctica
without BSA maintained PMI as well as C
+
and showed significant differences respect to the other treatments.
The percentage of cells thawed with protein extracts of
D. antarctica
and with cryoprotectants showed higher MMP than C
+
.
While, treatments without permeating and non-permeating cryoprotectants maintained a similar MMP to C
+
. AFPs from
D. antarctica
showed a cryoprotective effect in
S. salar
spermatozoa and these would act as non-permeating cryoprotectant,
replacing BSA in standard freezing medium.
Biography
Short S completed her Biotechnology Engineering at Universidad de La Frontera and started her Doctoral studies in Applied Cellular and Molecular Biology at
the Universidad de La Frontera. She is currently TA of Enzymology, Protein Structure and Immunology at the same institution. She has published five articles in
reputed journals.
s.short01@ufrontera.clShort S et al., J Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C2-092