Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation
Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation
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DOI：
10.1093/humrep/16.6.1191
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发表时间：
2001-06-01
影响因子：
6.1
通讯作者：
Lewis, SEM
中科院分区：
文献类型：
--
作者：
Donnelly, ET;Steele, EK;Lewis, SEM
人类精子的冷冻保存广泛用于人工授精和IVF计划。尽管在冷冻保存方法的各种进步，功能解冻后的精子的回收率仍然平庸，与精子活力显着下降冷冻后，本研究的目的是探讨冷冻保存对精子的DNA完整性和形态的影响从生育和不育的男人。精液样本来自17名生育男性和40名不育男性。通过不连续Percoll密度离心（95.0：47.5）制备所有样品。将样本分成等份，以便直接比较来自同一射精的新鲜和冷冻精子。将用于冷冻保存的等分试样与市售冷冻保护剂混合，并在投入液氮之前通过静态相蒸气冷却冷冻。在室温下缓慢解冻，使用改良的碱性单细胞凝胶电泳（彗星）试验测定精子DNA完整性，并使用Tygerberg标准分析精子形态。冷冻保存对正常生育男性精液和精子的DNA无明显影响，而不育男性精子则明显受损。冷冻保存对精液的形态和从生育和不育男性制备的样品有不利影响。
Cryopreservation of human spermatozoa is extensively used in artificial insemination and IVF programmes. Despite various advances in cryopreservation methodology, the recovery rate of functional post-thaw spermatozoa remains mediocre, with sperm motility being significantly decreased after freezing, This aim of this study was to investigate the effects of cryopreservation on both DNA integrity and morphology of spermatozoa from fertile and infertile men. Semen samples were obtained from 17 fertile and 40 infertile men. All samples were prepared by discontinuous Percoll density centrifugation (95.0:47.5). Samples were divided into aliquots to allow direct comparison of fresh and frozen spermatozoa from the same ejaculate. Aliquots for cryopreservation were mixed with a commercial cryoprotectant and frozen by static phase vapour cooling before plunging into liquid nitrogen. Thawing was carried out slowly at room temperature, Sperm DNA integrity was determined using a modified alkaline single cell gel electrophoresis (comet) assay and sperm morphology analysed using the Tygerberg criteria. DNA of semen and prepared spermatozoa from fertile men was found to be unaffected by cryopreservation, In marked contrast, spermatozoa from infertile men were significantly damaged by freeze-thawing. Cryopreservation had a detrimental effect on morphology of semen and prepared samples from fertile and infertile men.