OBJECTIVE: The aim of this experimental prospective study was to investigate the efficacy of single and combination sperm wash methods for their ability to isolate DNA intact spermatozoa. MATERIAL AND METHODS: Sperm DNA damage was introduced by local testicular irradiation in male mice and the extent of damage was quantified by comet assay. The spermatozoa were subjected to single (swim up or density gradient method) and also a combination of sperm wash techniques. The DNA integrity in various sub-fractions of wash techniques was evaluated. RESULTS: The amount of DNA damaged sperm did not differ between individual fractions when single wash technique was applied. However, a combination of density gradient and swim-up techniques significantly reduced (p<0.01) the number of DNA damaged sperm in the final population. CONCLUSION: The combination of density gradient separation and swim-up method is effective in eliminating DNA damaged spermatozoa.
OBJECTIVE: The aim of this experimental prospective study was to investigate the efficacy of single and combination sperm wash methods for their ability to isolate DNA intact spermatozoa. MATERIAL AND METHODS: Sperm DNA damage was introduced by local testicular irradiation in male mice and the extent of damage was quantified by comet assay. The spermatozoa were subjected to single (swim up or density gradient method) and also a combination of sperm wash techniques. The DNA integrity in various sub-fractions of wash techniques was evaluated. RESULTS: The amount of DNA damaged sperm did not differ between individual fractions when single wash technique was applied. However, a combination of density gradient and swim-up techniques significantly reduced (p<0.01) the number of DNA damaged sperm in the final population. CONCLUSION: The combination of density gradient separation and swim-up method is effective in eliminating DNA damaged spermatozoa.
Entities:
Keywords:
DNA damage; Sperm wash; comet assay; density gradient separation; swim-up
Authors: S K Adiga; M Toyoshima; K Shiraishi; T Shimura; J Takeda; M Taga; H Nagai; P Kumar; O Niwa Journal: Oncogene Date: 2007-04-09 Impact factor: 9.867