Bencharif D, Amirat L, Anton M, Schmitt E, Desherces S, Delhomme G, Langlois M-L, Barrière P, Larrat M, Tainturier D
Laboratory of Biotechnology and Reproductive Pathology, National Veterinary School of Nantes, BP 40706, 44307 Nantes, France.
Theriogenology. 2008 Dec;70(9):1478-88. doi: 10.1016/j.theriogenology.2008.06.095. Epub 2008 Sep 24.
A medium containing LDL (Low Density Lipoproteins, the cryoprotective component of chicken egg yolk) was compared with egg yolk for the preservation canine spermatozoa during the freeze-thaw process. Twenty sperm samples taken from 10 dogs were frozen in liquid nitrogen at -196 degrees C in seven different media: one control medium containing 20% egg yolk, and six test media containing 4%, 5%, 6%, 7%, 8%, and 10% LDL, respectively. Following thawing, sperm motility was assessed using a Hamilton-Thorne Sperm Analyser equipped with the CEROS 12 software. The percentage of motile spermatozoa was 55.3% in the 6% LDL medium (optimal concentration) compared with 27.7% in the egg yolk based medium (p<0.05). In comparison with the egg-yolk medium, the LDL medium also resulted in an improved preservation of spermatozoa during the freezing process (p<0.05) in terms of acrosomal integrity (FITC-PSA test), flagellar plasma membrane integrity (HOS test), and DNA integrity (Acridine Orange test). In addition, six Beagle bitches were inseminated twice, via the intra-uterine route, at an interval of 24h; 200x10(6) spermatozoa that had been previously frozen in the 6% LDL medium were used per insemination. All of the bitches became pregnant (gestation rate of 100%). In conclusion, the 6% LDL medium provides improved protection of the spermatozoa during the freeze-thaw process and a marked improvement in the motility parameters of canine spermatozoa in comparison with the control medium containing egg yolk alone. Finally, the use of LDL as a cryoprotectant for canine semen does not interfere with fertility.
在冷冻 - 解冻过程中,将含有低密度脂蛋白(LDL,鸡卵黄的冷冻保护成分)的培养基与卵黄用于犬精子的保存进行比较。从10只犬采集的20份精子样本在7种不同培养基中于 -196℃液氮中冷冻:一种对照培养基含20%卵黄,六种试验培养基分别含4%、5%、6%、7%、8%和10%的LDL。解冻后,使用配备CEROS 12软件的汉密尔顿 - 索恩精子分析仪评估精子活力。6%LDL培养基(最佳浓度)中活动精子的百分比为55.3%,而基于卵黄的培养基中为27.7%(p<0.05)。与卵黄培养基相比,LDL培养基在冷冻过程中还能更好地保存精子(p<0.05),在顶体完整性(FITC - PSA试验)、鞭毛质膜完整性(HOS试验)和DNA完整性(吖啶橙试验)方面均有改善。此外,6只比格犬母犬通过子宫内途径每隔24小时授精两次;每次授精使用先前在6%LDL培养基中冷冻的200×10⁶精子。所有母犬均怀孕(妊娠率100%)。总之,与仅含卵黄的对照培养基相比,6%LDL培养基在冷冻 - 解冻过程中能更好地保护精子,显著改善犬精子的活力参数。最后,使用LDL作为犬精液的冷冻保护剂不会影响生育能力。