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Published on: September 1, 2009
Frequent structural chromosome aberrations in immotile human sperm exposed to culture media
1Department of Anatomy, Hirosaki University School of Medicine, 5 Zaifucho, Hirosaki 036-8562, Japan. sage@cc.hirosaki-u.ac.jp
Background:
The influence of culture media or centrifugation on chromosomes of immotile human sperm was examined using ICSI into mouse oocytes.
Methods:
In experiment 1, immotile and motile human sperm retrieved directly from ejaculates were injected into mouse oocytes. In experiment 2, immotile human sperm were exposed to seminal plasma or one of four kinds of culture media (HEPES-BWW, modified-BWW, modified-human tubal fluid (HTF) and Dulbecco's phosphate-buffered saline) for 1.5-2.5 h at 18 degrees C in air before microinjection. In experiment 3, immotile human sperm were centrifuged along with HEPES-BWW before microinjection. In experiment 4, frozen-thawed immotile human sperm washed with seminal plasma or HEPES-BWW were injected into mouse oocytes. The hybrid oocytes were prepared for chromosome slides at first cleavage metaphase and were then examined cytogenetically.
Results:
In experiment 1, there was no significant difference in the incidences of structural chromosome aberrations between motile and immotile sperm (4.3% versus 5.8%). In experiment 2, culture media caused more frequent structural chromosome aberrations (14.3-32.6%) in immotile sperm than did seminal plasma (5.4%). In experiment 3, structural chromosome aberrations were found in 48.1% of the centrifuged immotile sperm, and a live/dead sperm viability test intimated that the aberrant sperm were probably dead. In experiment 4, the incidence of structural chromosome aberrations in frozen-thawed immotile sperm was significantly higher in HEPES-BWW (62.2%) than in seminal plasma (17.2%).
Conclusions:
The results indicate that immotile sperm do not have significantly more DNA lesions than motile sperm, although DNA of immotile sperm appears to be vulnerable to damage caused by different culture media.
Insights
Immotile sperm have similar DNA integrity to motile sperm. However, exposure to certain culture media or centrifugation can damage immotile sperm chromosomes.
Area of Science:
- Reproductive biology and genetics
- Spermatozoa chromosome integrity
- Assisted reproductive technologies
Background:
- Investigating the chromosomal status of immotile human sperm is crucial for understanding their suitability for assisted reproductive technologies.
- Previous research has not fully elucidated the impact of laboratory procedures on the genetic material of immotile sperm.
Purpose of the Study:
- To assess the influence of different culture media and centrifugation on the chromosomal integrity of immotile human sperm.
- To compare the frequency of chromosome aberrations in immotile sperm versus motile sperm.
- To evaluate the impact of cryopreservation and washing protocols on immotile sperm chromosomes.
Main Methods:
- Intracytoplasmic sperm injection (ICSI) of immotile and motile human sperm into mouse oocytes.
- Exposure of immotile sperm to various culture media (HEPES-BWW, modified-BWW, modified-HTF, Dulbecco's PBS) or seminal plasma before ICSI.
- Centrifugation of immotile sperm with HEPES-BWW prior to ICSI and cytogenetic examination of resulting metaphase chromosomes.
Main Results:
- No significant difference in chromosome aberrations between motile (4.3%) and immotile sperm (5.8%) directly from ejaculates.
- Culture media significantly increased chromosome aberrations in immotile sperm (14.3–32.6%) compared to seminal plasma (5.4%).
- Centrifugation (48.1%) and cryopreservation with HEPES-BWW (62.2%) led to higher rates of chromosomal damage in immotile sperm compared to seminal plasma (17.2%).
Conclusions:
- Immotile sperm do not inherently possess more DNA lesions than motile sperm.
- The DNA of immotile sperm is susceptible to damage induced by specific culture media and laboratory processing techniques.
- Careful selection of media and handling procedures is essential when utilizing immotile sperm in assisted reproduction.
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