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Evaluation of different fragment sizes and cryoprotectants for cryopreservation of feline testicular tissues
Author(s) -
Macente BI,
Toniollo GH,
Apparicio M,
Mansano CFM,
Thomé HE,
Canella CL,
Tozato MEG,
Gutierrez RR
Publication year - 2017
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12828
Subject(s) - cryoprotectant , cryopreservation , andrology , fragment (logic) , biology , microbiology and biotechnology , medicine , embryo , computer science , algorithm
Contents This study aimed to evaluate tissue damage of feline testicles sectioned in two different sizes (0.3 or 0.5 cm 3 ) and submitted to different cryoprotectants (propanediol or glycerol). Testicles obtained from 12 domestic cats were sectioned in 0.3 and 0.5 cm 3 sized pieces and immediately evaluated by TBARS and semi‐quantitatively by histomorphology. The remaining fragments were placed in cryotubes with 1 ml Egg yolk Tris Equex STM extender containing 3% glycerol or 3% propanediol and cryopreserved by fast‐freezing technique. Frozen‐thawed fragments were also evaluated by TBARS and histomorphology. Statistical analysis was performed using one‐way ANOVA with Student–Newmann–Keuls post hoc test, with p < .05. Fresh and cryopreserved tissues generally exhibited a similar morphology concerning detachment of cells from the basement membrane and observation of nucleoli, with a great proportion scored as 0 (no alteration). When present, alterations were slight and the morphology was considered to be good (most classified in scores 1). Pyknosis was the main anomaly observed as score 2 in 54.6% and 58.4% of 0.3‐cm 3 fragments cryopreserved in propanediol and glycerol, respectively (16.7% scored 2 in fresh tissue). In TBARS evaluation, 0.5‐cm 3 fragments cryopreserved in glycerol produced less free radical compared to the 0.3 cm 3 cryopreserved in glycerol or propanediol. Our results showed that glycerol was more efficient than propanediol to cryopreserve 0.5‐cm 3 fragments; this might be attributed to the fact that glycerol molecular weight is larger than propanediol and so its perfusion in the testicular tissue is slower.