Somatic Cell Nuclear Transfer (SCNT) in Mammals

The Cytoplast and Its Reprogramming Activities
  • Josef FulkaJr.
  • Helena Fulka
Part of the Advances in Experimental Medicine and Biology book series (volume 591)


It is now more than nine years since Dolly, the world’s first somatic cell cloned mammal was born, and the success of somatic cell nuclear transfer (SCNT) is still disappointingly low. Only about 3–5% of reconstructed embryos develop to term, and it is also evident that even if some clones are born, they are not necessarily fully developed and healthy. Embryonic and neonatal abnormalities of cloned offspring are probably a result of incorrect or incomplete reprogramming of the transferred donor cell nuclei. Such an incomplete reprogramming reflects the extremely low efficiency of SCNT. The key role in the process of reprogramming has been attributed to the enucleated oocyte-cytoplast into which the somatic cell nucleus is transferred. In our chapter, we will discuss the methodological approaches used for the preparation of cytoplasts and their possible reprogramming activities.


Polar Body Somatic Cell Nuclear Transfer Germinal Vesicle Mouse Oocyte Immature Oocyte 


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  1. 1.
    Hochedlinger K, Jaenisch R. Nuclear transplantation, embryonic stem cells, and the potential for cell therapy. N Engl J Med 2003; 349(3):275–286.PubMedCrossRefGoogle Scholar
  2. 2.
    Fulka Jr J, First NL, Moor RM. Nuclear and cytoplasmic determinants involved in the regulation of mammalian oocyte maturation. Mol Hum Reprod 1998; 4(1):41–49.PubMedCrossRefGoogle Scholar
  3. 3.
    Moor RM, Dai Y, Lee C et al. Oocyte maturation and embryonic failure. Hum Reprod Update 1998; 4(3):223–236.PubMedCrossRefGoogle Scholar
  4. 4.
    Hiragi T, Solter D. Reprogramming is essential in nuclear transfer. Mol Reprod Dev 2005; 70(4):417–421.CrossRefGoogle Scholar
  5. 5.
    Li E. Chromatin modification and epigenetic reprogramming in mammalian development. Nature Rev Genet 2002; 3(9):662–673.CrossRefGoogle Scholar
  6. 6.
    Khorasanizadeh S. The nucleosome: From genomic organization to genomic regulation. Cell 2004; 116(2):259–272.PubMedCrossRefGoogle Scholar
  7. 7.
    Fulka Jr J, Martinez F, Tepla O et al. Somatic and embryonic cell nucleus transfer into intact and enucleated immature mouse oocytes. Hum Reprod 2002; 17(8):2160–2164.PubMedCrossRefGoogle Scholar
  8. 8.
    Byrne JA, Simonsson S, Western PS et al. Nuclei of adult mammalian somatic cells are directly reprogrammed to oct-4 stem cell gene expression by amphibian oocytes. Curr Biol 2003; 13(14):1206–1213.PubMedCrossRefGoogle Scholar
  9. 9.
    Christians E, Boiani M, Garagna S et al. Gene expression and chromatin organization during mouse development. Dev Biol 1999; 207(1):76–85.PubMedCrossRefGoogle Scholar
  10. 10.
    Kim JM, Liu H, Tazaki M et al. Changes in histone acetylation during mouse oocyte meiosis. J Cell Biol 2003; 162(1):37–46.PubMedCrossRefGoogle Scholar
  11. 11.
    Fulka Jr J, Loi P, Ledda S et al. Nucleus transfer in mammals: How the oocyte cytoplasm modifies the transferred nucleus. Theriogenology 2001; 55(6): 1373–1380.PubMedCrossRefGoogle Scholar
  12. 12.
    Gao S, Gasparrini B, McGarry et al. Germinal vesicle material is essential for nucleus remodeling after nuclear transfer. Biol Reprod 2002; 67(3):928–934.PubMedCrossRefGoogle Scholar
  13. 13.
    Fulka Jr J, Loi P, Fulka H et al. Nucleus transfer in mammals: Noninvasive approaches for the preparation of cytoplasts. Trends Biotechnol 2004; 22(6):279–283.PubMedCrossRefGoogle Scholar
  14. 14.
    Li GP, White KL, Bunch TD. Review of enucleation methods and procedures used in animal cloning: State of the art. Cloning Stem Cells 2004; 6(1):5–13.PubMedCrossRefGoogle Scholar
  15. 15.
    Kim TM, Hwang WS, Shin JH et al. Development of nonmechanical enucleation method using X-ray irradiation in somatic cell nuclear transfer. Fertil Steril 2004; 82(4):963–965.PubMedCrossRefGoogle Scholar
  16. 16.
    Fulka Jr J, Moor RM. Noninvasive chemical enucleation of mouse oocytes. Mol Reprod Dev 1993; 34(4):427–430.PubMedCrossRefGoogle Scholar
  17. 17.
    Fisher Russell D, Ibanez E, Albertini DF et al. Activated bovine cytoplasts prepared by demecolcine-induced enucleation support development of nuclear transfer embryos in vitro. Mol Reprod Dev 2005; 72(2):161–170.CrossRefGoogle Scholar
  18. 18.
    Santos F, Dean W. Epigenetic reprogramming during early development in mammals. Reproduction 2004; 127(6):643–651.PubMedCrossRefGoogle Scholar
  19. 19.
    Beaujean N, Taylor J, Gardner J et al. Effect of limited DNA methylation reprogramming in the normal sheep embryo on somatic cell nuclear transfer. Biol Reprod 2004; 71(1): 185–193.PubMedCrossRefGoogle Scholar
  20. 20.
    Bourchis D, Le Bourhis D, Patin D et al. Delayed and incomplete reprogramming of chromosome methylation patterns in bovine cloned embryos. Curr Biol 2001; 11(19):1542–1546.CrossRefGoogle Scholar
  21. 21.
    Shi W, Haaf T. Aberrant methylation pattern at the two-cell stage as an indicator of early developmental failure. Mol Reprod Dev 2002; 63(3):329–334.PubMedCrossRefGoogle Scholar
  22. 22.
    Campbell KHS, Alberio R. Reprogramming the genome: Role of the cell cycle. Reproduction 2003; 61(Suppl):477–494.PubMedGoogle Scholar
  23. 23.
    Peterson CL, Laniel MA. Histones and histone modifications. Curr Biol 2004; 14(14):R546–551.PubMedCrossRefGoogle Scholar
  24. 24.
    Santos F, Peters AH, Otte AP et al. Dynamic chromatin modifications characterise the first cell cycle in mouse embryos. Dev Biol 2005; 280(1):225–236.PubMedCrossRefGoogle Scholar
  25. 25.
    Morgan HD, Santos F, Green K et al. Epigenetic reprogramming in mammals. Hum Mol Genet 2005; 14 (Rev Issue 1):R47–R58.PubMedCrossRefGoogle Scholar
  26. 26.
    Lepikhov K, Walter J. Differential dynamics of histone H3 methylation at positions K4 and K9 in the mouse zygote. BMC Dev Biol 2004; 4(12):1–5.Google Scholar

Copyright information

© Landes Bioscience and Springer Science+Business Media 2007

Authors and Affiliations

  • Josef FulkaJr.
    • 1
  • Helena Fulka
    • 1
    • 2
  1. 1.Institute of Animal ProductionPrague 10Czech Republic
  2. 2.Institute of Experimental MedicineCzech Academy of SciencesPragueCzech Republic

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