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Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification
Author(s) -
Pradiee J.,
Esteso M. C.,
Castaño C.,
ToledanoDíaz A.,
LopezSebastián A.,
Guerra R.,
SantiagoMoreno J.
Publication year - 2017
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12629
Subject(s) - cryoprotectant , extender , sperm , cryopreservation , glycerol , andrology , sperm motility , vitrification , yolk , sucrose , acrosome , chemistry , semen , dimethyl sulfoxide , biology , food science , biochemistry , embryo , medicine , organic chemistry , polyurethane , microbiology and biotechnology
Summary This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre‐freezing equilibration times (2–3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 m m ). The highest quality ( p < .01) thawed spermatozoa were obtained when using the TEST extender and an equilibration time of 3 hr. Sperm motility and membrane integrity were strongly reduced when using rapid freezing rates (60–85°C min −1 ), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 m m ) provided the highest ( p < .05) percentage of motile spermatozoa and live spermatozoa with an intact acrosome. Vitrified–warmed sperm variables were at their best when the spermatozoa was diluted in TCG–6% egg yolk + 100 m m sucrose and warmed at 60°C. Slow warming at 37°C strongly reduced ( p < .05) sperm motility and viability. However, sperm vitrification returned lower fertility, sperm motility and sperm viability values than conventional sperm freezing.