Advertisement

Microinjection pp 361-371 | Cite as

Electrofusion of 2-Cell Embryos for Porcine Tetraploid Embryo Production

  • Qingran Kong
  • Zhonghua Liu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1874)

Abstract

The electrofusion of 2-cell embryos proves to be a simple and efficient way of generating mammalian tetraploid embryos. Many factors affect the fusion efficiency, such as fusion medium, electric field intensity, and fusion pulse length. In mice, production of tetraploid embryos by electrofusion has already been investigated; however, the investigation to produce porcine tetraploid embryos is seldom reported. In this chapter, we will describe oocytes in vitro maturation, in vitro fertilization, and the optimum conditions for electrofusion of 2-cell embryos to produce tetraploid embryos in pig.

Key words

Pig Tetraploid embryo Electrofusion 

References

  1. 1.
    Luo YL, Lin L, Bolund L et al (2012) Genetically modified pigs for biomedical research. J Inherit Metab Dis 35:695–713CrossRefGoogle Scholar
  2. 2.
    Yang XZ, Smith SL, Tian XC et al (2007) Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning. Nat Genet 39:295–302CrossRefGoogle Scholar
  3. 3.
    Eakin GS, Hadjantonakis AK, Papaioannou VE et al (2005) Developmental potential and behavior of tetraploid cells in the mouse embryo. Dev Biol 288:150–159CrossRefGoogle Scholar
  4. 4.
    Jouneau A, Zhou Q, Camus A et al (2006) Developmental abnormalities of NT mouse embryos appear early after implantation. Development 133:1597–1607CrossRefGoogle Scholar
  5. 5.
    Zhao XY, Lv Z, Li W et al (2010) Production of mice using iPS cells and tetraploid complementation. Nat Protoc 5:963–971CrossRefGoogle Scholar
  6. 6.
    Kang L, Gao S (2012) Pluripotency of induced pluripotent stem cells. J Anim Sci Biotechnol 3:5CrossRefGoogle Scholar
  7. 7.
    Eakin GS, Behringer RR (2003) Tetraploid development in the mouse. Dev Dyn 228:751–766CrossRefGoogle Scholar
  8. 8.
    Modlinski JA (1978) Transfer of embryonic nuclei to fertilised mouse eggs and development of tetraploid blastocysts. Nature 273:466–467CrossRefGoogle Scholar
  9. 9.
    Sembon S, Fuchimoto D, Iwamoto M et al (2011) A simple method for producing tetraploid porcine parthenogenetic embryos. Theriogenology 76:598–606CrossRefGoogle Scholar
  10. 10.
    Bai C, Liu H, Liu Y et al (2011) Diploid oocyte formation and tetraploid embryo development induced by cytochalasin B in bovine. Cell Reprogram 13:37–45CrossRefGoogle Scholar
  11. 11.
    Eglitis MA (1980) Formation of tetraploid mouse blastocysts following blastomere fusion with polyethylene glycol. J Exp Zool 213:309–313CrossRefGoogle Scholar
  12. 12.
    Krivokharchenko A, Galat V, Ganten D, Bader M (2002) In vitro formation of tetraploid rat blastocysts after fusion of two-cell embryos. Mol Reprod Dev 61:460–465CrossRefGoogle Scholar
  13. 13.
    O’neill GT, Speirs S, Kaufman MH (1990) Sex-chromosome constitution of postimplantation tetraploid mouse embryos. Cytogenet Cell Genet 53:191–195CrossRefGoogle Scholar
  14. 14.
    Kaufman MH, Webb S (1990) Postimplantation development of tetraploid mouse embryos produced by electrofusion. Development 110:1121–1132PubMedGoogle Scholar
  15. 15.
    Ozil JP, Modlinski JA (1986) Effects of electric field on fusion rate and survival of 2-cell rabbit embryos. J Embryol Exp 96:211–228Google Scholar
  16. 16.
    Iwasaki S, Kono T, Fukatsu H, Nakahara T (1989) Production of bovine tetraploid embryos by electrofusion and their developmental capability in vitro. Gamete Res 24:261–267CrossRefGoogle Scholar
  17. 17.
    Prather RS, Hoffman KE, Schoenbeck RA, Stumpf TT, Li J (1996) Characterization of DNA synthesis during the 2-cell stage and the production of tetraploid chimeric pig embryos. Mol Reprod Dev 45:38–42CrossRefGoogle Scholar
  18. 18.
    Prochazka R, Vodicka P, Zudova D et al (2004) Development of in vivo derived diploid and tetraploid pig embryos in a modified medium NCSU 37. Theriogenology 62:155–164CrossRefGoogle Scholar
  19. 19.
    Hiriart MI, Bevacqua RJ, Canel NG et al (2013) Production of chimeric embryos by aggregation of bovine egfp eight-cell stage blastomeres with two-cell fused and asynchronic embryos. Theriogenology 80:357–364CrossRefGoogle Scholar
  20. 20.
    Kubiak JZ, Tarkowski AK (1985) Electrofusion of mouse blastomeres. Exp Cell Res 157:561–566CrossRefGoogle Scholar
  21. 21.
    Sawicki WAJ, Blaton O (1978) DNA synthesis in the second and third cell cycles of mouse preimplantation development. A cytophotometric study. Exp Cell Res 112:7CrossRefGoogle Scholar
  22. 22.
    Liu Z, Song J, Wang Z et al (2008) Green fluorescent protein (GFP) transgenic pig produced by somatic cell nuclear transfer. Sci Bull 53:1035–1039CrossRefGoogle Scholar
  23. 23.
    He W, Kong Q, Shi Y et al (2013) Generation and developmental characteristics of porcine tetraploid embryos and tetraploid/diploid chimeric embryos. Genomics Proteomics Bioinformatics 11:327–333CrossRefGoogle Scholar
  24. 24.
    Suo L, Wang F, Zhou GB, Shi JM, Wang YB, Zeng SM, Tian JH, Zhu SE, Liu GS (2009) Optimal concentration of calcium and electric field levels improve tetraploid embryo production by electrofusion in mice. J Reprod Dev 55:383–385CrossRefGoogle Scholar
  25. 25.
    Clement-Sengewald A, Brem G (1989) Electrofusion parameters for mouse two-cell embryos. Theriogenology 32:159–169CrossRefGoogle Scholar
  26. 26.
    Cheong HT, Taniguchi T, Hishinuma M, Takahashi Y, Kanagawa H (1991) Effects of various electric fields on the fusion and in vitro development of mouse two-cell embryos. Theriogenology 36:875–885CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Qingran Kong
    • 1
  • Zhonghua Liu
    • 1
  1. 1.Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life ScienceNortheast Agricultural UniversityHarbinChina

Personalised recommendations