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直接上游法与不连续Percoll梯度离心法联合用于少弱精子症样本的精子制备

Combination of direct swim-up technique and discontinuous Percoll gradient centrifugation for sperm preparation of oligoasthenozoospermic samples.

作者信息

Chen S U, Ho H N, Chen H F, Chao K H, Wu M Y, Chen C D, Huang S C, Lee T Y, Yang Y S

机构信息

Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Republic of China.

出版信息

Arch Androl. 1996 Sep-Oct;37(2):103-9. doi: 10.3109/01485019608988510.

Abstract

Sperm recovery for assisted reproduction in oligoasthenozoospermic patients is not satisfying either by the swim-up technique or by Percoll gradient centrifugation, and no single technique is constantly preferred. The design of this study was to evaluate the effects of combining the two methods on improving the efficacy of sperm preparation in these poor samples. For each semen sample, 1 mL was treated with a combination method, which used direct swim-up technique to recover motile sperm swimming to the supernatant, and then the residual semen was subjected to two-layer discontinuous Percoll gradient procedure for further recovery of motile sperm. Another 1 mL was prepared with two-layer discontinuous Percoll gradient centrifugation alone for comparison. Parameters measured included sperm concentration, number of progressively motile sperm, percentage of progressive motility, percentage of motile sperm recovery, amount of debris, percentage of normal forms according to Kruger's strict criteria, and motion characteristics of sperm using computer-aided motility analysis. The results of 30 oligoasthenozoospermic samples demonstrated that the combination method achieved a significantly greater recovery of motile sperm than the two-layer discontinuous Percoll gradient centrifugation did (43.2 +/- 19.7% vs. 32.2 +/- 14.3%, p < .05). The percentage of progressive motility was higher in the samples of the combination method than in those of Percoll gradient centrifugation alone, but the difference was not significant (63.7 +/- 21.8% vs. 58.7 +/- 20.1%). The debris of semen was removed equally well by both methods. The percentage of normal forms as well as motion characteristics, including curvilinear velocity, straight-line velocity, mean amplitude of lateral head displacement, and linearity, were similar in the samples treated by these two procedures. The combination of the direct swim-up technique and discontinuous Percoll gradient centrifugation combines the advantages of each procedure and results in greater recovery of motile sperm in oligoasthenozoospermic samples.

摘要

对于少弱精子症患者,无论是采用上游法还是Percoll梯度离心法进行辅助生殖的精子回收效果都不尽人意,且没有一种技术始终受到青睐。本研究的设计目的是评估将这两种方法结合起来对提高这些质量较差样本中精子制备效果的影响。对于每份精液样本,取1 mL采用联合方法处理,该方法先用直接上游法回收游向上清液的活动精子,然后将剩余精液进行两层不连续Percoll梯度离心程序以进一步回收活动精子。另取1 mL仅采用两层不连续Percoll梯度离心法制备作为对照。测量的参数包括精子浓度、进行性活动精子数量、进行性活动百分比、活动精子回收率、碎片量、根据克鲁格严格标准的正常形态百分比以及使用计算机辅助活动分析的精子活动特征。30份少弱精子症样本的结果表明,联合方法获得的活动精子回收率显著高于两层不连续Percoll梯度离心法(43.2±19.7%对32.2±14.3%,p<0.05)。联合方法样本中的进行性活动百分比高于仅采用Percoll梯度离心法的样本,但差异不显著(63.7±21.8%对58.7±20.1%)。两种方法去除精液碎片的效果相同。这两种程序处理的样本中,正常形态百分比以及活动特征,包括曲线速度、直线速度、头部侧向平均位移幅度和线性度都相似。直接上游法和不连续Percoll梯度离心法相结合兼具了每种程序的优点,能使少弱精子症样本中活动精子的回收率更高。

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