Quality assurance of human sperm for ICSI, exclusion of sperm with double and single strand breaks

1.本研究建立了单细胞脉冲场电泳（SPCPGE）技术，并利用单细胞脉冲场电泳技术中的摆动平台，将DNA纤维延长至0.5- 1.0mm。2.人精子低氧培养抑制了DNA纤维双链断裂的诱导。其中，核碱基类似物可能影响精子发生，尤其是对精子DNA的损伤。本文建立了用平衡沉降法、差速离心法和上游法完全去除洗涤后的精子中HIV的方法。SPCFGE结果显示，精子悬液中几乎所有精子的DNA均未发生双链断裂。4.人精浆中含有胞外DNA片段。本文建立了一种用酸变性法从DNA纤维中选择性提取腺嘌呤的方法。DNA含量与精子密度、活力、形态等精液质量无相关性。在Klieinfelter综合征中非常高。

1.The present study developed single cell pulse field electrophoresis (SPCPGE) to develop whole DNA fibers, furthermore they were elongated up to 0.5-1.0 mm by the use of pendular motion stage in SPCFGE.2.Human sperm culture in low oxygen concentration suppressed the induction of double strand breaks in DNA fibers.3.The patients with HIV usually take anti-HIV drugs. In these, nuclear base analogs might affect spermatogenesis, especially some damages might occur in sperm DNA. The present study established the method for complete exclusion of HIV from the washed sperm suspension by means of equilibrium sedimentation, differential velocity centrifugation and subsequent swim up. SPCFGE confirmed that almost all the sperm in the resulting sperm suspension were without double strand break in DNA.4.Human seminal plasma involved extracellular DNA fragments. The present study developed the method for selective adenine extraction from DNA fibers by means of acid denature. The amount of DNA was not correlated with semen qualities such as sperm concentration, motility, morphology. It was extremely high in Klieinfelter syndrome.