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- Title
Effects of sperm preparation on sperm DNA fragmentation and morphology.
- Authors
Hashimoto, S.; Goda, S.; Akamatsu, Y.; Yamanaka, M.; Morimoto, Y.
- Abstract
Introduction: Sperm DNA quality has been revealed to affect embryogenesis after fertilization. However, effect of sperm preparation on sperm DNA fragmentation is not well understood. In this study, the effect of sperm preparation on sperm DNA fragmentation was assessed using the DNA fragmentation index (DFI) and high DNA stainability (HDS) using flow cytometric analysis. Materials/Methods: Ejaculates containing at least 2.0 x 107 spermatozoa/ml and showing at least 50% progressive sperm motility were obtained from healthy men. Informed consent was obtained from each donor. The spermatozoa were prepared using a discontinuous gradient and/or a swim-up method. Samples were collected from original semen, sperm pellet centrifuged with HTF at 370 g for 10 min (A) and swim-up (B), or sperm pellet centrifuged with a discontinuous gradient at 620 g for 30 min (C) and swim-up (D). Each sample was assessed sperm quality based on the Kruger's strict criteria and was examined sperm DFI and HDS using sperm chromatin structure assay (SCSA, Evenson et al., 2000). Ultra-structures of spermatozoa treated with both a discontinuous gradient and a swim-up method and of original spermatozoa were assessed using TEM. Results: DFI of swim-up treated sample B (0.2--0.1%) and D (0.1%) were significantly lower (P < 0.05) than those of original (6.3--4.8%), sample A (3.9--3.6%) and C (2.0--1.4%). %) ) HDS rates of swim-up treated sample B (0.3--0.2%) and D (0.2%) were also significantly lower (P < 0.05) than those of original (2.2--2.4%), sample A (2.4--2.6%) and C (0.9--1.3%). The proportion of abnormal spermatozoa in sample C and D treated with a discontinuous gradient were lower than those in sample A and original. More morphological aberrations in original spermatozoa were observed by TEM compared with spermatozoa treated with both a discontinuous gradient and a swim-up. Conclusion: The present results indicate that centrifugation using a discontinuous gradient eliminates morphologically abnormal spermatozoa and swim-up treatment decreases DFI and HDS of spermatozoa. It is also shown that sperm preparation using both a discontinuous gradient and a swim-up improves sperm quality. Thus, it is inferred that sperm preparation using both a discontinuous gradient and a swim-up is effective for improvement of embryo development.
- Subjects
SPERMATOZOA; DNA
- Publication
Reproductive BioMedicine Online (Reproductive Healthcare Limited), 2008, Vol 16, Issue S2, pS-28
- ISSN
1472-6483
- Publication type
Abstract
- DOI
10.1016/S1472-6483(10)61532-5