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  • 1 Agricultural Biotechnology Centre H-2100 Gödöllő, P.O. Box 411, Hungary
  • 2 Agricultural Biotechnology Centre H-2100 Gödöllő, P.O. Box 411, Hungary
  • 3 Department and Clinic for Obstetrics and Reproduction, Faculty of Veterinary Sciences, Szent István University H-1400 Budapest, P.O. Box 2, Hungary
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This article presents a new, simple and rapid embryo biopsy method. The blastomere for genetic analysis can be separated from a precompacted mouse embryo after a partial zona digestion with the use of a holding pipette. For the micromanipulation only two microcapillaries and micromanipulators are needed. The development of the biopsied embryos was studied during in vitro culture and in utero following embryo transfer. There was no significant difference between the treated and the control groups in the ratio of embryos that developed to the blastocyst stage, although the biopsied embryos were delayed in their development because they contained significantly fewer cells compared to the control ones at the same stage. Although there was no difference in the ratio of implantation, the development of the biopsied embryos in utero was also delayed 12-24 hours on the 9th day of pregnancy. No difference in development was visible from the 13th day of pregnancy. Statistically, no differences were found in the developmental ratio (number of developed fetuses/transferred embryos) of the control and treated embryos during gastrulation (9th day of pregnancy), at the beginning of organogenesis (13th day of pregnancy) and before birth (19th day of pregnancy). The embryo biopsy method presented here can be a new and useful tool for preimplantation genetic diagnosis.

  • Hsu, M. I., Barroso, G., Mayer, J., Lanzendorf, S., Gibbons, W. E., Muasher, S. and Oehninger, S. 2000: Is the timing of implantation affected by zona pellucida micromanipulation?. J. Assist. Reprod. Genet. 17, 34-38.

    'Is the timing of implantation affected by zona pellucida micromanipulation?. ' () 17 J. Assist. Reprod. Genet. : 34 -38.

    • Search Google Scholar
  • Kaufman, M. H. 1990: Morphological stages of postimplantational embryonic development. In: Copp, A. J. and Cockroft, D. L. (eds) Postimplantational Mammalian Embryos. A Practical Approach. Oxford University, Oxford Press. pp. 81-91.

    Morphological stages of postimplantational embryonic development , () 81 -91.

  • Kaufman, M. H. 1992: The Atlas of Mouse Development. Academic Press Limited, London. pp. 47-67, 145-175.

    The Atlas of Mouse Development , () 47 -67.

  • Krzyminska, U. B., Lutjen, J. and O'Neill, C. 1990: Assessment of the viability and pregnancy potential of mouse embryos biopsied at different preimplantation stages of development. Hum. Reprod. 5, 203-208.

    'Assessment of the viability and pregnancy potential of mouse embryos biopsied at different preimplantation stages of development. ' () 5 Hum. Reprod. : 203 -208.

    • Search Google Scholar
  • Malter, H. E. and Cohen, J. 1989: Blastocyst formation and hatching in vitro following zona drilling of mouse and human embryos. Gamete Res. 24, 67-80.

    'Blastocyst formation and hatching in vitro following zona drilling of mouse and human embryos. ' () 24 Gamete Res. : 67 -80.

    • Search Google Scholar
  • Santalo, J., Grossmann, M. and Egozcue, J. 1996: Does Ca2+/Mg2+-free medium have an effect on the survival of the preimplantation mouse embryo after biopsy?. Hum. Reprod. Update 2, 257-261.

    'Does Ca2+/Mg2+-free medium have an effect on the survival of the preimplantation mouse embryo after biopsy?. ' () 2 Hum. Reprod. Update : 257 -261.

    • Search Google Scholar
  • Van Blerk, M., Nijs, M. and Van Steirteghem, A. C. 1991: Decompaction and biopsy of late mouse morulae: assessment of in-vitro and in-vivo developmental potential. Hum. Reprod. 6, 1298-1304.

    'Decompaction and biopsy of late mouse morulae: assessment of in-vitro and in-vivo developmental potential. ' () 6 Hum. Reprod. : 1298 -1304.

    • Search Google Scholar
  • Chen, S. U., Chao, K. H., Wu, M. Y., Chen, C. D., Ho, H. N. and Yang, Y. S. 1998: The simplified two-pipette technique is more efficient than the conventional three-pipette method for blastomere biopsy in human embryos. Fertil. Steril. 69, 569-575.

    'The simplified two-pipette technique is more efficient than the conventional three-pipette method for blastomere biopsy in human embryos. ' () 69 Fertil. Steril. : 569 -575.

    • Search Google Scholar
  • Critser, E. S. and First, N. L. 1986: Use of a fluorescent stain for visualization of nuclear material in living oocytes and early embryos. Stain. Technol. 61, 1-5.

    'Use of a fluorescent stain for visualization of nuclear material in living oocytes and early embryos. ' () 61 Stain. Technol. : 1 -5.

    • Search Google Scholar
  • Cui, K. H., Pannall, P., Cates, G. and Matthews, C. D. 1993: Blood analysis of mice born following single-cell embryo biopsy. Hum. Reprod. 8, 1906-1909.

    'Blood analysis of mice born following single-cell embryo biopsy. ' () 8 Hum. Reprod. : 1906 -1909.

    • Search Google Scholar
  • Cui, K. H., Barua, R. and Matthews, C. D. 1994: Histopathological analysis of mice born following single cell embryo biopsy. Hum. Reprod. 9, 1146-1152.

    'Histopathological analysis of mice born following single cell embryo biopsy. ' () 9 Hum. Reprod. : 1146 -1152.

    • Search Google Scholar
  • Wilton, L. J. and Trounson, A. O. 1989: Biopsy of preimplantation mouse embryos: development of micromanipulated embryos and proliferation of single blastomeres in vitro. Biol. Reprod. 40, 145-152.

    'Biopsy of preimplantation mouse embryos: development of micromanipulated embryos and proliferation of single blastomeres in vitro. ' () 40 Biol. Reprod. : 145 -152.

    • Search Google Scholar
  • Chen, S. U., Ho, H. N., Chen, H. F., Chao, K. H., Huang, S. C., Lee, T. Y. and Yang, Y. S. 1997: A simplified technique for embryo biopsy: use of the same micropipette for zona drilling and blastomere aspiration. J. Assist. Reprod. Genet. 14, 157-161.

    'A simplified technique for embryo biopsy: use of the same micropipette for zona drilling and blastomere aspiration. ' () 14 J. Assist. Reprod. Genet. : 157 -161.

    • Search Google Scholar
  • Dumoulin, J. C., Bras, M., Coonen, E., Dreesen, J., Geraedts, J. P. and Evers, J. L. 1998: Effect of Ca2+/Mg2+-free medium on the biopsy procedure for preimplantation genetic diagnosis and further development of human embryos. Hum. Reprod. 13, 2880-2883.

    'Effect of Ca2+/Mg2+-free medium on the biopsy procedure for preimplantation genetic diagnosis and further development of human embryos. ' () 13 Hum. Reprod. : 2880 -2883.

    • Search Google Scholar
  • Gimenez, C., Egozcue, J. and Vidal, F. 1994: Sexing sibling mouse blastomeres by polymerase chain reaction and fluorescent in-situ hybridization. Hum. Reprod. 9, 2145-2149.

    'Sexing sibling mouse blastomeres by polymerase chain reaction and fluorescent in-situ hybridization. ' () 9 Hum. Reprod. : 2145 -2149.

    • Search Google Scholar
  • Gordon, J. W. and Gang, I. 1990: Use of zona drilling for safe and effective biopsy of murine oocytes and embryos. Biol. Reprod. 42, 869-876.

    'Use of zona drilling for safe and effective biopsy of murine oocytes and embryos. ' () 42 Biol. Reprod. : 869 -876.

    • Search Google Scholar
  • Greenlee, A. R., Krisher, R. L. and Plotka, E. D. 1998: Rapid sexing of murine preimplantation embryos using a nested, multiplex polymerase chain reaction (PCR). Mol. Reprod. Dev. 49, 261-267.

    'Rapid sexing of murine preimplantation embryos using a nested, multiplex polymerase chain reaction (PCR). ' () 49 Mol. Reprod. Dev. : 261 -267.

    • Search Google Scholar
  • Handyside, A. H. and Delhanty, J. D. 1997: Preimplantation genetic diagnosis: strategies and surprises. Trends Genet. 13, 270-275.

    'Preimplantation genetic diagnosis: strategies and surprises. ' () 13 Trends Genet. : 270 -275.

    • Search Google Scholar
  • Hogan, B., Beddington, R., Constantini, F. and Lacy, E. 1994: Manipulating of mouse embryo. Second edition. Cold Spring Harbor Laboratory Press, New York.

    Manipulating of mouse embryo. Second edition , ().

  • Bodó, Sz., Baranyai, B., Gócza, E., Dohy, J. and Markkula, M. 2001: Preimplantation genetic diagnosis in cattle. Acta Vet. Hung. 49, 99-109.

    'Preimplantation genetic diagnosis in cattle. ' () 49 Acta Vet. Hung. : 99 -109.

  • Bredbacka, P. 2001: Progress on methods of gene detection in preimplantation embryos. Theriogenology 55, 23-34.

    'Progress on methods of gene detection in preimplantation embryos. ' () 55 Theriogenology : 23 -34.

    • Search Google Scholar