Cryopreservation of canine spermatozoa using a skim milk‐based extender and a short equilibration time
Cryopreservation of canine spermatozoa using a skim milk‐based extender and a short equilibration time
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使用脱脂牛奶补充剂和短平衡时间冷冻保存犬精子
DOI：
10.1111/rda.13806
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发表时间：
2020
影响因子：
1.7
通讯作者：
Suzuki Hiroshi
中科院分区：
文献类型：
--
作者：
Abe Yasuyuki;Asano Tomoyoshi;Wakasa Ichiko;Kume Aiko;Yokozawa Sakimi;Umemiya‐Shirafuji Rika;Suzuki Hiroshi
虽然已经建立了有用的精子冷冻保存技术，但在冷冻许多物种（包括犬）的精子之前，似乎需要长期平衡。本研究评价了5只雄性犬的冷冻保存犬精子在含棉子糖的脱脂乳（SM）基补充剂中缩短平衡期（0、30、60、120和180 min）后的生育力。当精子在室温（RT）下用稀释液稀释并冷冻保存而不平衡时，解冻后总活动精子（TMS）的比例（27%）低于平衡30 min（33%），60 min（32%），120 min（44%;p<0.05）或180 min（29%）时的比例。TMS的比例随着平衡时间的增加而增加，并在120 min时达到峰值。未经历初始平衡的冷冻保存精子的顶体完整性显著低于平衡的精子（p<0.05）。顶体正常率随着第一次平衡时间的延长而增加，并在120分钟时达到峰值。当冷冻解冻的精子已在室温下稀释，并进行了持续60或180分钟的初始平衡经宫颈授精到受体，有没有差异的交付率，窝仔数或繁殖效率。在使用SM-基稀释液冷冻保存犬精子时，即使初始平衡时间缩短至60 min，结果也与使用常规方法（初始平衡时间为180 min）时获得的结果相当。
Although useful spermatozoa cryopreservation techniques have been established, long‐term equilibration seems to be required before freezing the spermatozoa of many species, including dogs. The fertility of cryopreserved dog spermatozoa from five males for a reduced equilibration period (0, 30, 60, 120 and 180 min) in a skim milk (SM)‐based extender containing raffinose was evaluated in the present study. When the sperm was diluted with the extender at room temperature (RT) and cryopreserved without equilibration, the proportion of total motile spermatozoa (TMS) after thawing was lower (27%) than when the sperm was equilibrated for 30 min (33%), 60 min (32%), 120 min (44%;p< .05) or 180 min (29%). The proportion of TMS increased as the equilibration time increased and peaked at 120 min. Acrosome integrity was significantly lower in the cryopreserved spermatozoa that had not undergone the initial equilibration than in the equilibrated spermatozoa (p< .05). The normal rate of acrosomes increased with the extension of the first equilibration and peaked at 120 min. When frozen–thawed spermatozoa that had been diluted at RT and subjected to an initial equilibration lasting 60 or 180 min were transcervically inseminated into recipients, there were no differences in the delivery rate, litter size or breeding efficiency. In the cryopreservation of canine spermatozoa using a SM‐based extender, even if the initial equilibration time was shortened to 60 min, the results were comparable to those obtained when the conventional method (with an initial equilibration time of 180 min) was used.