A new paper by our group with the first authorship of Zoran Marinović was accepted for publication in
General and Comparative Endocrinology as a part of the proceedings of last year's
5th International Workshop on the Biology of Fish Gametes held in Ancona, Italy on September 7-11, 2015. For more information please consult this link:
http://www.sciencedirect.com/science/article/pii/S0016648016302027
Cryosurvival
of isolated testicular cells and testicular tissue of tench Tinca tinca
and goldfish Carassius auratus following slow-rate freezing
Abstract
Experiments were
carried out to test the efficiency of cryopreservation of whole testicular
tissue in tench Tinca tinca and goldfish Carassius auratus and compare it to
cryopreservation of isolated testicular cells. Additionally, effects of three
cryoprotectants (dimethyl sulphoxyde – Me2SO, methanol – MeOH and ethylene
glycol – EG) at three concentrations (1 M, 2 M and 3 M) on post-thaw cell
viability were assessed. Tissue pieces / isolated testicular cells were diluted
in cryomedia and cryopreserved by slow-rate freezing (1 °C/min to – 80 °C
followed by a plunge into the liquid nitrogen). In both species Me2SO and EG
generally yielded higher cryosurvival of early-stage germ cells than MeOH,
while spermatozoa of neither species displayed such a pattern. In most cases a
3M > 2M > 1M viability pattern emerged in both species for both sample
types regardless of the cryoprotectant used. Sample type (dissociated
testicular cells vs testicular tissue) did not seem to affect viability rates
of tench early-stage germ cells and goldfish spermatozoa, while the opposite
was observed for tench spermatozoa and goldfish early-stage germ cells.
Additionally, through histological analysis we displayed that tissue structure
mainly remained unaltered after thawing in goldfish. These results indicate
that cryopreservation of whole testicular tissue is indeed a valid alternative
method to cryopreservation of dissociated testicular cells. Early-stage germ
cells obtained from cryopreserved testis can be further used in different
purposes such as transplantation into suitable donors while viable sperm might
be used for fertilization when feasible.
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