两种精子优选法对精子活率、尾部低渗肿胀率和形态正常精子百分率的影响

Effects of two sperm selection methods on sperm viability,hypoosmotic swelling rate of human spermatozoa tail and percentage of human normal morphology spermatozoa

目的:评价Percoll密度梯度离心和上游精子分离方法对人精子活率、尾部低渗肿胀率和形态正常精子百分率影响。方法:收集12例男性不育症患者精液样本,分别采用Percoll密度梯度离心和上游精子分离方法(简称上游法)进行精子优选,应用伊红-Y染色分析优选前后精子活率和尾部低渗肿胀率,应用改良巴氏染色法分析优选前后精子形态。结果:①Percoll法和上游法优选后精子活率和尾部低渗肿胀率均明显高于优选前(P<0.01),两种优选法间比较精子活率和尾部低渗肿胀率差异均无显著性(P>0.05)。Percoll法和上游法优选后B-G型精子尾部低渗肿胀率均明显高于优选前(P<0.01),两种优选法间比较B-G型精子尾部低渗肿胀率差异均无显著性(P>0.05)。②Percoll密度梯度离心法和上游法优选后形态正常精子百分率均明显高于优选前(P<0.05),上游法优选后形态正常精子百分率明显高于Percoll法优选后形态正常精子百分率(P<0.05)。上游法优选后锥型头、无定型头和颈/中段缺陷形态异常精子百分率均明显少于优选前(P<0.05);Percoll法优选后锥型头、无定型头和尾部缺陷形态异常精子百分率均明显少于优选前(P<0.05);上游法优选后无定型头、颈/中段缺陷和尾部缺陷形态异常精子百分率明显低于Percoll优选后(P<0.05)。结论:Percoll法和上游法优选后精子活率、尾部低渗肿胀率和形态正常精子百分率较优选前明显改善,此两种优选法均可广泛应用于生殖医学,其中上游法优于Percoll法。

Objective To study the effects of Percoll gradient centrifugation and swimming up technique on the sperm viability, the hypoosmotic swelling rate of human spermatozoa tail and the percentage of morphologically normal spermatozoa. Methods Semen samples were collected from 12 male infertility patients. Sperm selection was performed respectively using Percoll gradient centrifugation and swimming up technique. Sperm viabilities and the hypoosmotic swelling rates of human spermatozoa tail before and after sperm selection were analyzed by Eosin stain. Sperm morphological changes before and after sperm selection were evaluated by Modified Papanicolaou stain. Results ①The sperm viability and the hypoosmotic swelling rate of human spermatozoa tail were significantly increased after Percoll gradient centrifugation and swimming up （P〈0.01）. No significant difference was found in comparison of sperm viability and the hypoosmotic swelling rate of human spermatozoa tail between the two selection methods （P〉0.05）. The rate of sperm with type B-G tail swelling was significantly improved after Percoll and swimming up （P〈0.01）, However, there was no significant difference of type B-G tail swelling rate between the two selection methods. ②The rate of morphologically normal sperm significantly was increased after Percoll and swimming up, Meanwhile, the rate of normal morphological sperm after swimming up was higher than that after percoll （P〈 0.05）. The rate of Tapered sperm, Irregularity sperm and Neck/Midpiece defect sperm after swimming up were significantly decreased （P〈0.05）. The rate of Tapered sperm, Irregularity sperm, and Tail defect sperm after Percoll were significantly decreased. However, the rate of Irregularity sperm, Neck/Midpiece defect sperm and tail defect sperm after swimming up were significantly lower than those after Percoll （P〈0.05）. Conclusion After sperm selection using Percoll gradient centrifugation and swimming up technique, the sperm viability, hypoosmotic swelling rate of human spermatozoa tail and rate of morphologically normal spermatozoa are obviously improved. These two sperm selection methods may be widely applied in reproductive medicine. Swimming up technique is better than Percoll gradient centrifugation.