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  • Motility and fertility of rabbit sperm cryopreserved using soybean lecithin as an alternative to egg yolk.

Motility and fertility of rabbit sperm cryopreserved using soybean lecithin as an alternative to egg yolk.

Theriogenology (2015-07-26)
Kazutoshi Nishijima, Shuji Kitajima, Chihiro Koshimoto, Masatoshi Morimoto, Teruo Watanabe, Jianglin Fan, Yukihisa Matsuda
ABSTRACT

This study was conducted to investigate whether soy lecithin can be used as an alternative cryoprotectant to establish a procedure that does not require the use of egg yolk to cryopreserve rabbit strains. Semen from Japanese White rabbits was frozen with HEPES extender containing 20% egg yolk (EYH), 0.5% (Lec-0.5), 1.5% (Lec-1.5), 2.5% (Lec-2.5), or 3.5% (Lec-3.5; wt/vol) lecithin (type IV-S, ≥30%), and the motility of thawed sperm was analyzed. The sperm motility in the Lec-1.5 group was significantly higher than that in the Lec-2.5 and 3.5 groups and equivalent to the EYH group. From 17 rounds of artificial insemination with frozen-thawed sperm in the EYH and Lec-1.5 groups, 12 rabbits in both groups were pregnant (70.6%) and delivered offspring. The litter size was 3.3 in the EYH group and 5.1 in the Lec-1.5 group. These results indicate that soy lecithin can be used as a substitute for egg yolk as a cryoprotectant on the basis of motility and fertility of the frozen-thawed rabbit sperm and that 1.5% lecithin (type IV-S, ≥30%) in the semen extender was the optimum concentration for rabbit sperm cryopreservation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
HEPES, BioUltra, Molecular Biology, ≥99.5% (T)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)
Sigma-Aldrich
Acetamide, sublimed, 99%
Sigma-Aldrich
Acetamide, ~99% (GC)
Sigma-Aldrich
HEPES, Vetec, reagent grade, 99.5%
Sigma-Aldrich
HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
Sigma-Aldrich
HEPES buffer solution, 1 M in H2O