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

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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.