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Assisted fertilization of mouse oocytes and preliminary results for human oocytes using zona drilling

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Abstract

The zona-drilling procedure was investigated in mouse oocytes prior to a study on human oocytes. The procedure involved the injection of 5-nl volumes of acidic Hepes-buffered medium at pH 2.5 using a microinjection instrument. Zona-drilled mouse oocytes had significantly higher rates of fertilization (60/99; 61%) than zona-intact oocytes (6/103; 6%) at an insemination concentration of 1×104 sperm/ml (P<0.001). The procedure did not induce parthenogenetic activation of oocytes and more than 97% of zygotes developed to the blastocyst stage. A similar rate of live progeny was observed when zona-drilled (38.0%) and control embryos (38.5%) were transferred to pseudopregnant recipients. Chromosome analyses were performed on zona-intact, zona-free, and zona-drilled oocytes inseminated with varying concentrations of sperm and analysed at the first cleavage division. Zonafree oocytes had high rates of polyploidy (≥40%) with varying insemination numbers but the zona-drilled oocytes did not reveal significant increases in the rate of polyploidy or aneuploidy when compared to controls. In the human studies, zona-drilled oocytes achieved higher rates of fertilization than zona-intact oocytes, with sperm numbers as low as 1×104/ml (6/8; 75%). Polyspermic fertilization was observed in 1/2 and 2/6 of fertilized oocytes inseminated with 1×105 and 1×104 sperm/ml, respectively. With the low sperm concentration 2/4 of those which were normally fertilized developed to healthy blastocysts. These studies suggest that the zona-drilling technique as described can be performed without apparent harm to oocytes and generate normal embryos. Micromanipulation by zona drilling should be explored further as a potential treatment mode for cases of oligol asthenospermic infertility.

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Authors and Affiliations

  1. PIVET Medical Centre, 166-168 Cambridge Street, 6007, Perth, Western Australia

    W. R. Edirisinghe & J. L. Yovich

  2. Murdoch University, 6150, Perth, Western Australia

    R. G. Wales & H. M. Chapman

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  1. W. R. Edirisinghe
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  2. R. G. Wales
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  3. H. M. Chapman
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  4. J. L. Yovich
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Edirisinghe, W.R., Wales, R.G., Chapman, H.M. et al. Assisted fertilization of mouse oocytes and preliminary results for human oocytes using zona drilling. J Assist Reprod Genet 8, 48–55 (1991). https://doi.org/10.1007/BF01131591

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  • DOI: https://doi.org/10.1007/BF01131591

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