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Hum. Reprod. Advance Access originally published online on January 21, 2005
Human Reproduction 2005 20(5):1244-1247; doi:10.1093/humrep/deh757
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions{at}oupjournals.org

A novel method for chromosome analysis of human sperm using enucleated mouse oocytes

Yasuyuki Araki1,2,3,4, Midori Yoshizawa1,2 and Yasuhisa Araki3

1 Science of Plant and Animal Production, United Graduated School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchuu-city, Tokyo, 183-8509, 2 Department of Animal Breeding and Reproduction, Faculty of Agriculture, Utsunomiya University, 350, Mine-machi, Utsunomiya, Tochigi, 321-8505 and 3 Institute for Advanced Reproductive Medical Technology, 909-21, Ishii, Fujimi, Seta-gun, Gunma, 371-0105, Japan

4 To whom correspondence should be addressed at: Institute for Advanced Reproductive Medical Technology, 909-21, Ishii, Fujimi, Setagun, Gunma, 371-0105, Japan. Email: yaaraki{at}nifty.com

BACKGROUND: Mouse oocytes can be used in conjunction with intracytoplasmic sperm injection (ICSI) as a technique to permit chromosomal analysis of human sperm. However, chromosomes derived both from the human sperm and the mouse oocyte appear simultaneously following ICSI. The present study focused on evaluating whether or not previously enucleated mouse oocytes are usable for the analysis of human sperm chromosomes. METHODS: The metaphase chromosome–spindle complex was removed from a mouse oocyte. Human sperm from a donor with proven fertility were injected into mouse enucleated oocytes or intact oocytes. The presence of pronuclei in the oocytes was confirmed ~7–11 h after ICSI, and the oocytes were then fixed so that the nuclei could be observed as chromosome samples at 15–16 h after ICSI. RESULTS: The formation rate of one pronucleus in enucleated oocytes after ICSI was 93.9% (186/198) while that of two pronuclei in intact oocytes after ICSI was 85.4% (88/103). The appearance rate of metaphase chromosomes of human sperm in the enucleated oocytes, 89.4% (160/179), was significantly higher than that in intact oocytes, 78.7% (74/94) (P=0.017). CONCLUSIONS: An efficient ICSI method using enucleated mouse oocytes was established to allow the visualization of the human sperm chromosome complement without the risk of confusion with mouse oocyte chromosomes.

Key words: chromosomes/enucleated mouse oocytes/human sperm/ICSI


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Hum. Reprod., March 1, 2008; 23(3): 627 - 634.
[Abstract] [Full Text] [PDF]


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