Brennan A P, Holden C A
Monash IVF, Richmond, Victoria, Australia.
Hum Reprod. 1995 Sep;10(9):2308-12. doi: 10.1093/oxfordjournals.humrep.a136291.
In this study, human spermatozoa obtained from donors (n = 15) with normal semen characteristics were cryopreserved in human sperm preservation medium, supplemented with the phosphodiesterase inhibitor pentoxifylline at concentrations of 0, 1, 3 and 10 mM. The effect of pentoxifylline on cryopreserved spermatozoa was determined by monitoring changes in sperm motility and acrosome morphology by labelling the spermatozoa with fluorescein-conjugated concanavalin A lectin. Cryoprotectant supplemented with 1 mM pentoxifylline was found to improve post-thaw progressive motility from 15.3 +/- 2.4 (control) to 23.1 +/- 3.8% (P < 0.01), and total motility from 27.4 +/- 3.3 (control) to 38.2 +/- 3.9% (P < 0.05) without reducing the percentage of spermatozoa with normal acrosomal regions, and so appears useful for cryopreservation purposes. The beneficial effects of 1 mM pentoxifylline on sperm motility were shown to be maintained post-thaw over a 6 h time course. Cryoprotectant supplemented with 3 mM pentoxifylline was found to improve only post-thaw progressive motility, from 15.3 +/- 2.4 (control) to 20.7 +/- 3.0% (P < 0.05). However, cryopreservation in the presence of 10 mM pentoxifylline was found to have a significantly (P < 0.01) detrimental effect on acrosome morphology post-thaw, reducing it from 29.0 +/- 2.0 (control) to 21.0 +/- 2.4% without affecting sperm motility. This suggests that assessment of the acrosomal region may indicate subtle deleterious effects of cryoprotectant supplements that cannot be determined from post-thaw motility assessments alone. These findings differ from previous studies in that a lower concentration of pentoxifylline (1 mM) was found to be optimal for cryopreservation purposes.
在本研究中,从精液特征正常的供体(n = 15)获取的人类精子,保存在添加了浓度为0、1、3和10 mM磷酸二酯酶抑制剂己酮可可碱的人类精子保存培养基中。通过用荧光素偶联的伴刀豆球蛋白A凝集素标记精子,监测精子活力和顶体形态的变化,以确定己酮可可碱对冷冻保存精子的影响。发现添加1 mM己酮可可碱的冷冻保护剂可将解冻后的前向运动率从15.3±2.4(对照组)提高到23.1±3.8%(P<0.01),总运动率从27.4±3.3(对照组)提高到38.2±3.9%(P<0.05),且不降低顶体区域正常的精子百分比,因此似乎对冷冻保存有用。1 mM己酮可可碱对精子活力的有益作用在解冻后的6小时内得以维持。发现添加3 mM己酮可可碱的冷冻保护剂仅能提高解冻后的前向运动率,从15.3±2.4(对照组)提高到20.7±3.0%(P<0.05)。然而,发现在10 mM己酮可可碱存在下进行冷冻保存对解冻后的顶体形态有显著(P<0.01)有害影响,将其从29.0±2.0(对照组)降低到21.0±2.4%,而不影响精子活力。这表明对顶体区域的评估可能表明冷冻保护剂补充剂的细微有害影响,而仅通过解冻后活力评估无法确定这些影响。这些发现与先前的研究不同,因为发现较低浓度的己酮可可碱(1 mM)对冷冻保存最为适宜。