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法(SPCPGE)，利用单细胞脉冲场电泳仪中的摆动平台，使DNA纤维延伸到0.5-1.0 mm。2.低氧条件下培养的人精子可抑制DNA纤维的双链断裂。3.HIV患者常服用抗HIV药物。其中，核碱基类似物可能会影响精子的发生，尤其是精子DNA的一些损伤。本研究建立了用平衡沉淀法、差速离心法和上游法从洗涤的精子悬液中完全排除HIV的方法。SPCFGE证实，在得到的精子悬液中，几乎所有的精子都没有DNA双链断裂。4.人类精浆涉及细胞外DNA片段。本研究建立了用酸变性方法从DNA纤维中选择性提取腺嘌呤的方法。DNA含量与精子密度、精子活力、精子形态等精液质量无关。在Klieinfelter综合征中，这一比例极高。

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.