Kim Sung Woo, Nho Eun Jee, Lee Joong Yeup, Jee Byung Chul
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea.
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea.
Clin Exp Reprod Med. 2019 Dec;46(4):147-151. doi: 10.5653/cerm.2019.00458. Epub 2019 Nov 19.
The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST).
Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern ("a"-"g") were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed.
The HOST examinations showed that >93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type "d", 98.67% in type "g", and 98.17% in type "f" spermatozoa.
We found that the type "d" spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.
本研究旨在根据低渗肿胀试验(HOST)确定的特定尾部肿胀模式,研究人类精子中的DNA片段化状态。
将来自21名健康供体的冷冻精液样本解冻,并采用上游技术制备用于胞浆内单精子注射。作为HOST程序的一部分,精液样本处理5分钟,然后使用Halosperm试剂盒进行精子染色质扩散试验。在单个精子水平评估DNA片段化状态(大晕圈、中晕圈、小晕圈、无晕圈或降解)和特定尾部肿胀模式(“a”-“g”)。共分析了42000个精子,并根据观察到的特定尾部肿胀模式评估无DNA片段化的精子百分比(以大晕圈或中晕圈为证)。
HOST检查显示,所有类型的精子中>93%没有DNA片段化。“d”型精子中无DNA片段化的精子百分比为100%,“g”型为98.67%,“f”型精子为98.17%。
我们发现“d”型精子没有DNA片段化,但其他类型的精子也显示出非常低的DNA片段化率。这一结果可能与通过密度梯度离心和上游技术对精子的处理有关。