Bwanga C O, de Braganca M M, Einarsson S, Rodriguez-Martinez H
Department of Obstetrics and Gynaecology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala.
Zentralbl Veterinarmed A. 1990 Oct;37(9):651-8. doi: 10.1111/j.1439-0442.1990.tb00958.x.
Split ejaculates from four boars were frozen with a programmable freezing machine, in mini- (0.25 ml) and maxi- (5 ml) plastic straws with an extender at either acidic (6.3) or alkaline (7.4) pH. Glycerol (3%) was used as cryoprotectant. The freezing of the semen was monitored by way of thermocouples placed in the straws. Post-thaw motility and acrosome integrity were evaluated; the latter using phase contrast microscopy, eosin-nigrosin stain and electron microscopy. Post-thaw sperm motility was significantly higher when semen was frozen in mini-straws than in maxi-straws. For the mini-straws, the motility was better when semen was exposed to an acidic environment during freezing, but this beneficial effect of the low extracellular pH was not evident when maxi-straws were thawed. The motility of the spermatozoa diminished significantly during the thermoresistance test (0 h and 2 h time) at 37 degrees C in a similar way for both straws and extracellular pH's. The freezing procedure, no matter the extracellular pH, did not cause major acrosomal damages, but significantly more normal apical ridges were present in the mini-straws than in the maxi-straws. This in vitro evaluation indicated that the freezing method employed was better for mini- than for maxi-straws since the freezing of the 5 ml volumes was not homogeneous, due to the large section area between the surface and the core of the straw.
将来自四头公猪的精液分成几份,使用程序控制的冷冻机进行冷冻,分别装入微型(0.25毫升)和大型(5毫升)塑料细管中,稀释液的pH值为酸性(6.3)或碱性(7.4)。使用3%的甘油作为冷冻保护剂。通过置于细管中的热电偶监测精液的冷冻过程。解冻后评估精子活力和顶体完整性;后者采用相差显微镜、伊红-黑色素染色和电子显微镜进行评估。当精液在微型细管中冷冻时,解冻后的精子活力显著高于在大型细管中冷冻的精液。对于微型细管,冷冻过程中精液处于酸性环境时活力更佳,但解冻大型细管时,细胞外低pH值的这种有益效果并不明显。在37摄氏度的耐热性测试(0小时和2小时)期间,两种细管以及不同细胞外pH值条件下,精子活力均以相似的方式显著下降。无论细胞外pH值如何,冷冻过程均未造成严重的顶体损伤,但微型细管中的正常顶嵴明显多于大型细管。这种体外评估表明,所采用的冷冻方法对微型细管的效果优于大型细管,因为5毫升体积的精液在大型细管中冷冻不均匀,这是由于细管表面与核心之间的截面积较大所致。