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与简单的精子洗涤相比,新鲜单峰骆驼精液的单层离心可提高精子质量和体外受精能力。

Single layer centrifugation of fresh dromedary camel semen improves sperm quality and in vitro fertilization capacity compared with simple sperm washing.

作者信息

Malo C, Crichton E G, Morrell J M, Pukazhenthi B S, Skidmore J A

机构信息

Camel Reproduction Center, Dubai, United Arab Emirates.

Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.

出版信息

Reprod Domest Anim. 2017 Dec;52(6):1097-1103. doi: 10.1111/rda.13036. Epub 2017 Aug 12.

Abstract

Single layer centrifugation (SLC) through a colloid is a tool for selecting viable mammalian spermatozoa but has not been used previously for fresh dromedary camel sperm. Semen from six camels (2 ejaculates/male) was diluted 1:5 (v:v) or 1:10 (v:v) in a Tris-citrate-fructose buffer for mechanical liquefaction by gentle pipetting. Following liquefaction, semen was processed either by SLC or by centrifugation without a colloid (control). Total and progressive motilities, CASA kinematics, vitality and acrosome integrity (eosin-nigrosin) and plasma membrane integrity (Hypo-osmotic swelling test; HOST), and fertilizing ability in a heterologous assay (zona-free goat oocytes) were evaluated. Both total (p = .003) and progressive motilities (p = .003) were higher in SLC-processed than in control semen samples, irrespective of dilution. Positive HOST values increased when using colloid in 1:5 (p = .001) and 1:10 dilution (p = .010). Colloid-selected sperm had higher penetration rates than controls (p < .001 and p = .02 for 1:5 and 1:10 dilutions, respectively). However, only the SLC sperm at 1:5 dilution showed higher percentages of pronuclear formation (p = .02) than controls. Dilution effect was only significant for total motility before in vitro fertilization, with higher values for the 1:5 dilution (p = .033). The recovery rates of motile sperm between dilutions were similar (26.1% vs 35.4%; p = .226). In conclusion, SLC is a promising tool for selecting functional dromedary camel sperm and warrants more research.

摘要

通过胶体进行单层离心(SLC)是一种筛选有活力的哺乳动物精子的方法,但此前尚未用于新鲜的单峰骆驼精子。从六只骆驼采集的精液(每只雄性2次射精)在柠檬酸盐 - 果糖 - 三羟甲基氨基甲烷缓冲液中按1:5(v:v)或1:10(v:v)稀释,通过轻柔移液进行机械液化。液化后,精液分别通过SLC或不使用胶体的离心法(对照)进行处理。评估了总活力和渐进性活力、计算机辅助精子分析(CASA)运动学、活力、顶体完整性(伊红 - 黑色素)和质膜完整性(低渗肿胀试验;HOST),以及在异源试验(无透明带山羊卵母细胞)中的受精能力。无论稀释度如何,经SLC处理的精液样本的总活力(p = 0.003)和渐进性活力(p = 0.003)均高于对照精液样本。使用1:5(p = 0.001)和1:10稀释度(p = 0.010)的胶体时,HOST阳性值增加。经胶体筛选的精子穿透率高于对照(1:5和1:10稀释度分别为p < 0.001和p = 0.02)。然而,只有1:5稀释度的SLC精子显示出比对照更高的原核形成百分比(p = 0.02)。稀释效应仅在体外受精前对总活力有显著影响,1:5稀释度的值更高(p = 0.033)。不同稀释度之间活动精子的回收率相似(26.1%对35.4%;p = 0.226)。总之,SLC是一种筛选功能性单峰骆驼精子的有前途的工具,值得进一步研究。

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