封闭式薄片法在睾丸单精子冷冻保存中的应用探索

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摘要目的探讨封闭式薄片法对梗阻性无精子症（OA）患者睾丸单精子的冷冻效果,以期为非梗阻性无精子症（NOA）患者单精子冷冻保存提供一种简单有效的方式。方法选取2016年6~12月在大连市生殖与遗传医学中心诊断为OA 15例患者作为研究对象,应用封闭式薄片法冷冻方式对睾丸单精子进行冷冻保存。睾丸冷冻精子解冻复苏后（冷冻组）,对体外成熟（IVM）卵子行卵胞浆内单精子显微注射技术（ICSI）,统计其受精率与卵裂率,并与睾丸新鲜精子（新鲜组）ICSI进行比较,以期对睾丸单精子解冻后受精能力进行评估。冷冻组及新鲜组精子均来自上述15例患者。结果应用封闭式薄片法对睾丸单精子进行冷冻,解冻后具有很高的精子回收率与存活率（92.00%,65.94%）。冷冻组与新鲜组具有相似的受精率（56.00% vs. 70.66%）和卵裂率（97.44% vs. 95.83%）,两组差异无统计学意义（P>0.05）。结论封闭式薄片法玻璃化冷冻方式可有效保存OA患者睾丸单精子,且精子复苏后具有较高的受精能力,为NOA患者单精子冷冻保存应用于临床奠定了实验基础。

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	马超
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关键词 ：睾丸单精子, 冷冻保存, 无精子症, 回收率

Abstract：Objective To investigate the effect of Closed-Sheet Method on testicular spermatozoa in patients with obstructive azoospermia(OA),in order to provide a simple and effective cryopreservation for individual numbers of testicular spermatozoa in patients with non-obstructive azoospermia(NOA).Methods 15 patients were diagnosed with OA by Reproductive Medicine & Genetic Center of Dalian from June to December 2016.Single testicular spermatozoa was frozen with Closed-Sheet Method,and we compared cleavage rate and fertility rate after ICSI between frozen-thawed individual numbers of testicular spermatozoa and fresh testicular sperm with abandoned in vitro maturation of immature oocytes,and the fertilization rate of frozen-thawed testicular spermatozoa was evaluated.Both frozen and fresh groups were from the above 15 patients.Results The frozen-thawed individual numbers of testicular spermatozoa had higher recovery rate and viability rate(92.00%,65.94%) with the Closed-Sheet Method.The fertility rate (56.00% vs 70.66%)and cleavage rate (97.44% vs 95.83%) of frozen testicular spermatozoa was similar to fresh testicular sperm after ICSI,and no significant difference(P>0.05).Conclusion Since the frozen-thawed testicular spermatozoa had a higher ability of fertilization.The Closed-Sheet Method was effective to cryopreserve individual numbers of testicular spermatozoa in patients with OA.This study laid the experimental basis for the clinical application of single sperm cryopreservation in patients with NOA.

Key words： Individual numbers testicular spermatozoa Cryopreservation Azoospermia Recovery rate

收稿日期: 2017-06-27

PACS:

R321.33

基金资助:国家自然科学基金项目（81571498）

作者简介: 马超（1985-）,男,硕士,助理研究员,主要从事生殖实验室工作

引用本文:

马超, 胡明广, 金波. 封闭式薄片法在睾丸单精子冷冻保存中的应用探索[J]. 中国当代医药, 2017, 24(24): 19-22.
MA Chao, HU Ming-guang, JIN Bo. Application of the Closed-Sheet Method for individual numbers of testicular spermatozoa cryopreservation. 中国当代医药, 2017, 24(24): 19-22.

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http://dangdaiyiyao.com/CN/ 或 http://dangdaiyiyao.com/CN/Y2017/V24/I24/19

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