Cryopreservation of individually selected sperm: methodology and case report of a clinical pregnancy.
Cryopreservation of individually selected sperm: methodology and case report of a clinical pregnancy.
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DOI：
10.1007/s10815-012-9733-y
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发表时间：
2012-05
影响因子：
3.1
通讯作者：
Falcone, Tommaso
中科院分区：
文献类型：
--
作者：
Desai, Nina;Goldberg, Jeffrey;Austin, Cynthia;Sabanegh, Edmund;Falcone, Tommaso
关键词：
为了描述一种从睾丸活检组织中冷冻单独分离的精子的新技术，在冷冻前对附睾液和少精子精液样本进行了活动精子的评估。用ICSI针分离运动或抽动的精子进行单个精子的冷冻保存。选定的精子被装载在高安全性吸管(单纯疱疹病毒；欧文科学；欧文，加利福尼亚州)，在~0.5g L液体中，以便于回收。还使用常规方法将样品冷冻在冷藏箱(100-1000ug L等份)中。在这两种冷冻技术中，样品都是缓慢冷却的。试验用卵黄缓冲液-甘油(欧文)作为冷冻保护剂。进行解冻试验以评估精子回收率和活动率。6名患有无精子症的男性接受了单一精子的冷冻，并在传统的冷冻箱中冷冻了等量的睾丸或附睾精子。此外，两名患有少精子症的男性也进行了个别精子的选择和冷冻。在所有8例患者中，解冻过程中~0.5U L精子液很容易从吸管中卸出。精子回收率为33%~100%。除一个样本外，所有样本的运动性都很明显。在6例中，精子用于成熟卵母细胞的胞浆内单精子注射。除1例外，其余均受精。在这项研究中，我们报告了第一例使用这种技术的临床妊娠。这次怀孕值得注意的是，在最初的睾丸准备中识别和选择的单个活动精子被成功冷冻。我们随后能够回收这些精子，使一个卵母细胞受精，得到的胚胎产生了一个活生生的孩子。这份初步报告中描述的方法为隔离少量精子提供了一种新的方式。在精子发生严重受损的情况下，它可能特别有用，在这种情况下，可能需要进行广泛的筛查才能找到几个有活力的精子。
To describe a new technique for freezing individually isolated spermatazoa from testicular biopsies, epididymal aspirates and oligospermic semen samples Samples were evaluated for the presence of motile sperm before cryopreservation. Motile or twitching sperm were isolated with an ICSI needle for single sperm cryopreservation. Selected sperm were loaded on the High Security Straw (HSV; Irvine Scientific; Irvine,CA), in ~0.5 µl of fluid to facilitate recovery. The sample was also frozen using conventional methodology in cryovials (100–1000 µl aliquots). In both freezing techniques, the samples were slow cooled. Test-yolk buffer-glycerol (Irvine) was used as the cryoprotectant. Test-thaws were performed to assess sperm recovery and motility. Six men with azoospermia had single sperm cryopreservation, as well as freezing aliquots of their testicular or epididymal sperm in traditional cryovials. In addition, two men with oligospermia also had individual sperm selected and frozen. In all 8 cases, the ~0.5 µl of fluid containing sperm was quite easily unloaded from the HSV straw during thawing. The percent sperm recovery ranged from 33% to 100%. Motility was evident in all but one sample. In six cases, the sperm were used for intracytoplasmic sperm injection of mature oocytes. Fertilization occurred in all but one case. In this study, we report the first clinical pregnancy with this technique. This pregnancy was remarkable in that a single motile sperm identified and selected in the initial testicular preparation was successfully frozen. We were able to subsequently recover this sperm, fertilize an oocyte and the resultant embryo gave rise to a live birth. The methodology described in this preliminary report offers a new modality for sequestering small numbers of sperm. It may be particularly useful in cases involving severe impairment of spermatogenesis, where extensive screening may be necessary to find a few viable sperm.
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影响因子：
6.7
作者：
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通讯作者：
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