Sharma R K, Seifarth K, Agarwal A
Department of Urology, Cleveland Clinic Foundation, Ohio, USA.
Int J Fertil Womens Med. 1997 Nov-Dec;42(6):412-7.
Sperm recovery using a single-layer Percoll procedure is significantly better than using the swim-up technique for infertile men and patients with normal sperm characteristics; however, in normal men results have been contradictory. Some studies have shown further improvement in semen quality with multiple layers. Therefore, this study compared the effect of single-layer and two-layer Percoll procedures on sperm characteristics of normozoospermic men.
Semen specimens from 10 normal donors were processed by layering 1 mL of the liquefied ejaculate on a single layer of 80% Percoll or on a two-layer (47% and 90%) Percoll gradient. Computer-assisted semen analysis was done to examine total motile sperm, percentage of recovery of motile cells, percent motility, curvilinear velocity, linearity, and amplitude of lateral head displacement. Each specimen was evaluated by the hypo-osmotic swelling (HOS) test, bovine cervical mucus penetration test, viability (eosin-nigrosin stain), and sperm morphology (World Health Organization and Kruger's strict criteria).
Specimens processed with the two-layer Percoll procedure had significantly better recovery of spermatozoa, and significantly better percentage motility, linearity, amplitude of lateral head displacement, percentage tail swelling, and percentage viability than those separated on single-layer Percoll. Results for sperm morphology using WHO and Kruger's criteria were similar between the two methods (P = 0.92 for both sets of criteria).
In normozoospermic men, the two-layer Percoll separation procedure significantly improves semen characteristics compared with separation on a single layer.
对于不育男性和精子特征正常的患者，采用单层 Percoll 法回收精子明显优于上游法；然而，对于正常男性，结果却相互矛盾。一些研究表明，多层 Percoll 可进一步改善精液质量。因此，本研究比较了单层和两层 Percoll 法对正常精子男性精子特征的影响。
将 10 名正常供者的精液标本分别置于单层 80% Percoll 或两层（47%和 90%）Percoll 梯度上，加入 1 mL 液化精液进行处理。采用计算机辅助精液分析检测总活动精子、活动细胞回收率、活动率、曲线速度、直线性和头部侧摆幅度。每个标本通过低渗肿胀试验、牛宫颈黏液穿透试验、活力（伊红 - 黑色素染色）和精子形态（世界卫生组织和克鲁格严格标准）进行评估。
与单层 Percoll 分离的标本相比，两层 Percoll 法处理的标本精子回收率显著更高，活动率、直线性、头部侧摆幅度、尾部肿胀率和活力百分比也显著更高。两种方法根据世界卫生组织和克鲁格标准得出的精子形态结果相似（两组标准的 P 值均为 0.92）。
对于正常精子男性，两层 Percoll 分离法与单层分离相比，能显著改善精液特征。
