自制金属冷冻板法冷冻人附睾精子的实验研究  

Self-made metal freezing plate for cryopreservation of human epididymal sperm: An experimental study

作　　者：房芃[1] 赵秀红[1] 王洪霞 毕敬磊 陈永刚[1] 王爱妍 崔艳国[1] FANG Peng;ZHAO Xiu-hong;WANG Hong-xia;BI Jing-lei;CHEN Yong-gang;WANG Ai-yan;CUI Yan-guo(Center of Reproductive Medicine, Zibo Hospital of Maternity and Child Care, Zibo, Shandong 255000, China)

机构地区：[1]淄博市妇幼保健院生殖医学中心

出　　处：《中华男科学杂志》2019年第7期586-589,共4页National Journal of Andrology

摘　　要：目的:研究一种新型人精子冷冻方法对精子复苏率的影响,以探索人附睾精子、睾丸穿刺精子的最佳冷冻方法。方法:选取76例梗阻性无精子症患者的附睾穿刺(PESA)标本,按照自制金属冷冻板法和传统冷冻法分两组冷冻。采用计算机辅助精液分析系统检测冷冻前、后前向运动精子百分率,并对比两种方法对精子膜功能、DNA碎片指数(DFI)、顶体酶活性和精子畸形百分率的影响。结果:复苏后自制金属冷冻板法和传统冷冻法前向运动精子百分率[(12.0±7.5)%vs(8.0±5.1)%,P<0.05]和低渗肿胀精子百分率[(22.0±17.5)%vs(18.0±20.5)%]比较有显著性差异(P<0.05),均较冷冻前[(20.7±8.8)%和(30.0±13.5)%]显著下降(P<0.05)。自制金属冷冻板法复苏后精子顶体酶活性显著高于传统冷冻法[(75.2±9.5)μIU/10^6精子vs(55.7±8.3)μIU/10^6精子,P<0.05],均较冷冻前(120.0±10.5)μIU/10^6精子显著下降(P<0.05)。两种方法复苏后畸形精子百分率和DFI无显著差异[(98.7±8.8)%vs(98.5±9.2)%,P>0.05]和[(38.2±8.5)%vs(39.5±10.2)%,P>0.05],并均较冷冻前[(97.2±9.5)%和(30.8±9.7)%]显著升高(P<0.05)。自制金属冷冻板法和传统冷冻法冷冻复苏率[(65.2±12.0)%vs(52.3±18.0)%]有显著性差异(P<0.05)。结论:自制金属冷冻板法是一种经济高效、操作简单的精子冷冻方法且能最大限度的节约精子;复苏后可以保证较好的精子复苏率、活动力和顶体酶活性。Objective: To study the effect of a new human sperm freezing method on the sperm recovery rate and search for an optimal method for cryopreservation of human epididymal sperm. Methods: We collected semen samples from 76 men with obstructive azoospermia by percutaneous epididymal sperm aspiration and divided each sample into two parts to be cryopreserved with a self-made metal freezing plate(the experimental group) or by slow freezing(the control group), respectively. We measured the percentage of progressively motile sperm(PMS) with the computer-assisted semen analysis system and compared the membrane function, DNA fragmentation index(DFI), acrosin activity and morphological abnormality of the sperm between the two groups before and after cryopreservation. Results: After thawing, both the percentages of PMS and hypotonically swollen sperm were significantly higher in the experimental than in the control group([12.0 ± 7.5]% vs [8.0 ± 5.1]%, P < 0.05;[22.0 ± 17.5]% vs [18.0 ± 20.5]%, P < 0.05), though both decreased in comparison with the pre-freezing parameters([20.7 ± 8.8]% and [30.0 ± 13.5]%)(P < 0.05). The sperm acrosin activity was remarkably higher in the experimental than in the control group after thawing([75.2 ± 9.5] vs [55.7 ± 8.3]μIU/10^6 sperm, P < 0.05), though decreased as compared with the baseline([120.0 ± 10.5]μIU/10^6 sperm, P < 0.05). No statistically significant differences were observed between the experimental and the control groups after thawing in the percentage of morphologically abnormal sperm([98.7 ± 8.8]% vs [98.5±9.2]%, P > 0.05) or sperm DFI [38.2 ± 8.5]% vs [39.5 ± 10.2]%, P > 0.05), though both markedly elevated in comparison with the pre-freezing parameters([97.2 ± 9.5]% and [30.8 ± 9.7]%)(P < 0.05). The post-thaw recovery rate of sperm was significantly higher in the experimental than in the control group([65.2 ± 12.0]% vs [52.3 ± 18.0]%, P < 0.05). Conclusion: The self-made metal freezing plate, with its advantages of low cost, high efficiency, and easy operatio

关 键 词：金属冷冻板 精液冷冻 复苏率 附睾精子 辅助生殖技术 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]