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

1.本研究开发了单细胞脉冲场电泳（SPCPGE）来产生完整的DNA纤维，并通过SPCFGE中的摆动运动台将其延长至0.5-1.0毫米。2.低氧浓度下的人类精子培养抑制了DNA纤维双链断裂的诱导。3.HIV患者通常服用抗HIV药物。其中，核碱基类似物可能会影响精子发生，特别是精子 DNA 可能会发生一些损伤。本研究建立了通过平衡沉降、差速离心和随后的上游从洗涤的精子悬浮液中完全排除HIV的方法。 SPCFGE证实所得精子悬浮液中几乎所有精子的DNA均无双链断裂。4.人精浆含有细胞外DNA片段。本研究开发了通过酸变性从 DNA 纤维中选择性提取腺嘌呤的方法。 DNA 量与精子浓度、活力、形态等精液质量无关。 Klieinfelter 综合征中该值极高。

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.