Eradication of DNA damaged sperm using swim�up and density gradient separation methods

Dr. Ashwani Kumar Bansal; Dr. Neena Gupta; Dr Arpit Bansal; Dr. Vandana Bansal

Volume : VII, Issue : I, January - 2017

Eradication of DNA damaged sperm using swim�up and density gradient separation methods

Dr. Ashwani Kumar Bansal, Dr. Neena Gupta, Dr Arpit Bansal, Dr. Vandana Bansal

Abstract :

INTRODUCTION: Swim–up techniques for sperm separation may have detrimental effects on sperm DNA. We wished to determine whether the normal swim–up method with centrifugation used in our laboratory, which involves a centrifugation step, was harmful to sperm compared with swim–up without centrifugation. METHODS: Semen samples were obtained from patients undergoing IVF or andrology assessment. An aliquot was removed for fixation and subsequent DNA fragmentation determination. The remaining sample was divided into two equal parts, which was subjected to swim–up either with (normal swim–up) or without (direct–swim–up) centrifugation. Semen analysis was performed both before and after swim–up. DNA fragmentation, in spermatozoa previously fixed in 4% paraformaldehyde, was assessed by the terminal transferase–mediated DNA end labeling procedure (TUNEL). The percentage of spermatozoa with DNA damage after each swim–up technique was compared with that in the original semen sample. RESULTS: DNA damage was <5 % in most sample. No significant change in DNA fragmentation was observed between two swim–up procedures but significantly changes were observed between healthy men and infertile men. CONCLUSIONS: we conclude that our TUNEL technique based flowcytometry are useful to differentiate the live and damaged sperm with their percentage.