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Cryopreserved Immature Mouse Oocytes: A Chromosomal and Spindle Study

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Abstract

Purpose: Cryopreservation of human oocytes might provide an alternative approach to freezing supernumerary embryos obtained during IVF. This process, performed on immature denuded prophase I mouse oocytes, was investigated.

Methods: We first investigated the capacity of frozen, immature, murine oocytes to continue in vitro maturation after thawing. We then evaluated the risk to offspring from chromosomal damage by cytogenetical and cytological (spindle) analysis. Finally, we attempted to determine the reasons for and the stage of maturation failure.

Results: A total of 700 immature oocytes was frozen, 629 (90%) were recovered intact after thawing, and 53% extruded the first polar body, versus 74% for the control group. Freezing was not accompanied by an increase in aneuploidy in maturing oocytes (18 and 15% for thawed and control oocytes, respectively). Consequently, the first meiotic division occurred normally, without an increase in nondisjunction. Spindle analysis demonstrated only a few abnormalities (15 and 2% for thawed and control oocytes, respectively) incompatible with further development. Oocytes arrested during in vitro maturation were mainly at the metaphase I stage (64 and 76% for thawed and control oocytes, respectively). Whereas 17% of thawed oocytes were blocked before the formation of the first meiotic spindle, this never occurred in the control group.

Conclusions: Immature murine oocytes can withstand cryopreservation, which is encouraging for future human application of this technique.

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REFERENCES

  1. Glenister P, Wood M, Kirby C, Whittingham D: Incidence of chromosome anomalies in first-cleavage mouse embryos obtained from frozen-thawed oocytes fertilized in vitro. Gamete Res 1987;16:205–216

    Google Scholar 

  2. Pickering S, Johnson M: The influence of cooling on the organization of the meiotic spindle of the mouse oocyte. Hum Reprod 1987;3:207–216

    Google Scholar 

  3. Kola L, Kirby C, Shaw J, Davey A, Trounson A: Vitrification of mouse oocytes results in aneuploid zygotes and malformed fetuses. Teratology 1988;38:467–474

    Google Scholar 

  4. Bouquet M, Selva J, Auroux M: The incidence of chromosomal abnormalities in frozen-thawed mouse oocytes after in-vitro fertilization. Hum Reprod 1992;1:76–80

    Google Scholar 

  5. Toth T, Baka S, Veeck L, Jones H, Muasher S, Lanzendorf S: Fertilization and in vitro development of cryopreserved human prophase I oocytes. Fertil Steril 1994;5:891–894

    Google Scholar 

  6. Quinn P, Barros C, Whittingham G: Preservation of hamster oocytes to assay the fertilizating capacity of human spermatozoa. J Reprod Fertil 1982;66:161–168

    Google Scholar 

  7. Downs SM, Schroeder AC, Eppig JJ: Developmental capacity of mouse oocytes following maintenance of meiotic arrest in vitro. Gamete Res 1986;15:305–316

    Google Scholar 

  8. Candy CJ, Wood D, Whittingham G: Cryopreservation of immature mouse oocytes. Hum Reprod 1994;9:1738–1742

    Google Scholar 

  9. Maro B, Johnson MH, Pickering S: Changes in actin distribution during fertilization of the mouse egg. J Embryol Exp Morphol 1984;81:211–237

    Google Scholar 

  10. Kilmartin JV, Wright B, Milstein C: Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol 1982;93:576–582

    Google Scholar 

  11. Tarkowski AK: An air drying method for chromosome preparaion from mouse eggs. Cytogenetics 1966;5:394–400

    Google Scholar 

  12. Van Blerkom J: Maturation at high frequency of germinal vesicle-stage mouse oocytes after cryopreservation: Alterations in cytoplasmic, nuclear, nucleolar and chromosomal structure and organisation associated with vitrification. Hum Reprod 1989;4:883–898

    Google Scholar 

  13. Van Blerkom J, Davis P: Cytogenetic, cellular, and developmental consequences of cryopreservation of immature and mature mouse and human oocyte. Microsc Res Tech 1994;27:165–193

    Google Scholar 

  14. Van der Elst J, Nerinckx S, Van Sterteghem AC: In vitro maturation of germinal vesicle-stage oocytes following cooling, exposure to cryoprotectants and ultrarapid freezing: Limited effect on the morphology of the second meiotic spindle. Hum Reprod 1992;10:1440–1446

    Google Scholar 

  15. Van der Elst J, Nerinckx S, Van Steirteghem AC: Slow and ultrarapid freezing of fully grown germinal vesicle-stage mouse oocytes: Optimization of survival rate outweighed by defective blastcyst formation. J Assist Reprod Genet 1993;10:202–212

    Google Scholar 

  16. Gook D, Osborn M, Johnston W: Cryopreservation of mouse and human oocytes using 1,2-propanediol and the configuration of the meiotic spindle. Hum Reprod 1993;8:1101–1109

    Google Scholar 

  17. Ashwood-Smith M, Morris G, Fowler R, Appleton T: Physical factors are involved in the destruction of embryos and oocytes during freezing and thawing procedures. Hum Reprod 1988;3:795–802

    Google Scholar 

  18. Trounson A, Anderiesz C: The effect of testosterone on the maturation and developmental capacity of murine oocytes in vitro. Hum Reprod 1995;10:2377–2381

    Google Scholar 

  19. Mattson B, Albertini D: Oogenesis: Chromatin and microtubule dynamics during meiotic prophase. Mol Reprod Dev 1990;25:374–383

    Google Scholar 

  20. Veeck L, Baka S, Toth T, Jones H: Evaluation of the spindle apparatus of in-vitro matured human oocytes following cryopreservation. Hum Reprod 1995;7:1816–1820

    Google Scholar 

  21. Winston N, McGuinness O, Johnson M, Maro B: The exit of mouse oocytes from meiotic M-phase requires an intact spindle during intracellular calcium release. J Cell Sci 1995;103:389–396

    Google Scholar 

  22. Polanski Z: Activation of in vitro matured mouse oocytes arrested at the first or second metaphase. J. Dev Biol 1995;39:1015–1020

    Google Scholar 

  23. Eppig J, Schultz R, O'Brien M, Chesnel F: Relationship between the developmental programs controlling nuclear and cytoplasmic maturation of mouse oocytes. Dev Biol 1994;164:1–9

    Google Scholar 

  24. Bouquet M, Selva J, Auroux M: Cryopreservation of mouse oocytes: Mutagenic effects in the embryos? Biol Reprod 1993;49:764–769

    Google Scholar 

  25. Bouquet M, Selva J, Auroux M: Effects of cooling and equilibration in DMSO, and cryopreservation of mouse oocytes, on the rates of in vitro fertilization, development, and chromosomal abnormalities. Mol Reprod Dev 1995;40:110–115

    Google Scholar 

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Authors and Affiliations

  1. Reproductive Biology and Cytogenetics Laboratory, Hospital Kremlin Bicêtre, France

    N. Frydman, J. Selva, M. Bergere & M. Auroux

  2. Laboratoire de FIV, Unité du Pr Selva, Hôpital Antoine Béclère, 157 rue de la Porte de Trivaux, 92141, Clamart, France

    B. Maro

Authors
  1. N. Frydman
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  2. J. Selva
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  3. M. Bergere
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  4. M. Auroux
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  5. B. Maro
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Frydman, N., Selva, J., Bergere, M. et al. Cryopreserved Immature Mouse Oocytes: A Chromosomal and Spindle Study. J Assist Reprod Genet 14, 617–623 (1997). https://doi.org/10.1023/A:1022593004528

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