Department of Obstetrics and Gynaecology, University of Stellenbosch, Tygerberg, South Africa.
Andrologia. 2011 Jun;43(3):217-21. doi: 10.1111/j.1439-0272.2010.01136.x.
Sperm retrieval techniques form an integral part of the assisted reproductive programme. The success of sperm separation is measured by the number of motile sperm retrieved from a given semen sample. The study aimed to evaluate the effect of temperature during sperm preparation events on the number and percentage motile sperm retrieved following a double wash swim-up procedure. Thirty semen samples were obtained from 10 normozoospermic donors. After collection samples were divided into two aliquots, one aliquot was placed in an incubator at 34 °C, while the second aliquot was left at room temperature (25 °C). Sperm motility assessments were recorded with a computer assisted sperm analyser. Motile sperm fractions were retrieved from the semen samples following a double wash swim-up technique. Two tubes were prepared for each experiment. Tubes were placed in two different centrifuges: (i) SpermFuge (Shivani Industries, India) with temperature centrifuge control (34 °C) and (ii) Sigma with no temperature control facilities. Both centrifuges were set at 484 g for 5 min. Following the second wash, sperm pellets were layered with culture medium, and sperm was allowed to swim up. Supernatants were removed and analysed for sperm concentration and motility values. Percentage motile sperm was transformed to ARCSIN values and results of the two centrifugation methods at 34 °C and room temperature were compared with Mann-Whitney test for independent samples. The mean sperm concentration retrieved at 34 °C was 43.8 ± 50 (SpermFuge) and 32.7 ± 21 (Sigma) (P < 0.05), compared to retrieved concentration at room temperature namely, 30.9 ± 33 (SpermFuge) and 30.6 ± 17 (Sigma) (P ≥ 0.05). The mean percentage motile sperm at 34 °C was 64.0 ± 19 (SpermFuge) and 44.2 ± 24 (Sigma) (P = 0.02), while at room temperature the percentage motile sperm was 54.7 ± 17 (SpermFuge) compared to 46.5 ± 14 (Sigma) (P ≥ 0.05). Centrifuge temperature and incubation temperature significantly influenced the percentage retrieved motile sperm. The use of temperature-controlled sperm preparation might have clinical value for men with poor sperm motility values.
精子提取技术是辅助生殖计划的一个组成部分。精子分离的成功与否取决于从给定精液样本中提取的活动精子数量。本研究旨在评估精子制备过程中温度对双洗泳动后精子回收率和活力百分比的影响。从 10 名正常精子供体中获得 30 份精液样本。采集后,将样本分为两份,一份置于 34°C 的孵育箱中,另一份置于室温(25°C)下。使用计算机辅助精子分析仪记录精子活力评估。通过双洗泳动技术从精液样本中提取活动精子。每个实验准备了两个管。将两个管放入两个不同的离心机中:(i)具有温度离心机控制的 SpermFuge(Shivani Industries,印度)(34°C)和(ii)无温度控制设施的 Sigma。两个离心机均以 484 g 离心 5 分钟。第二次洗涤后,将精子沉淀层与培养基混合,让精子游动。移去上清液,分析精子浓度和活力值。将活动精子百分比转换为 ARCSIN 值,将 34°C 下两种离心方法的结果与独立样本的 Mann-Whitney 检验进行比较。34°C 下提取的精子浓度平均值为 43.8±50(SpermFuge)和 32.7±21(Sigma)(P<0.05),而室温下提取的浓度分别为 30.9±33(SpermFuge)和 30.6±17(Sigma)(P≥0.05)。34°C 下的平均活动精子百分比为 64.0±19(SpermFuge)和 44.2±24(Sigma)(P=0.02),而室温下的活动精子百分比为 54.7±17(SpermFuge),而 46.5±14(Sigma)(P≥0.05)。离心机温度和孵育温度显著影响了提取的活动精子百分比。对于精子活力值较低的男性,使用温度控制的精子制备可能具有临床价值。