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Related Experiment Videos

Reprogramming cattle somatic cells by isolated nuclear injection.

A Trounson1, O Lacham-Kaplan, M Diamente

  • 1Centre for Early Human Development, Institute of Reproduction and Development, Monash Universit, Monash Medical Centre, Clayton, Victoria, Australia.

Reproduction, Fertility, and Development
|December 28, 1999
PubMed
Summary
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Somatic cell nuclear transfer (SCNT) was optimized for producing cloned cattle embryos. Nuclear injection of somatic cell nuclei proved to be an efficient method for bovine embryo cloning.

Area of Science:

  • Reproductive Biology
  • Animal Biotechnology
  • Developmental Biology

Background:

  • Somatic cell nuclear transfer (SCNT) is a key technique for animal cloning.
  • Optimizing SCNT procedures is crucial for improving efficiency and developmental rates in cattle.

Purpose of the Study:

  • To describe the development of an SCNT procedure for producing blastocyst-stage cattle embryos.
  • To compare different SCNT methods, including electrofusion and nuclear injection, for their efficiency in cattle embryo cloning.

Main Methods:

  • Bovine fetal fibroblasts were used as donor cells.
  • Electrofusion parameters were optimized using isofusion contours.
  • Cytoplast size and double cytoplasts were evaluated for their effect on fusion and cleavage rates.

Related Experiment Videos

  • Comparison of subzonal perivitelline space injection, intracytoplasmic injection, and parthenogenetic activation.

    • Main Results:

    • Fusion rates increased with decreased cytoplast size, but cleavage rates decreased.
    • Development to blastocysts was not achieved using double cytoplasts.
    • Intracytoplasmic injection of nuclei resulted in 2% blastocyst development, while parthenogenetic activation yielded 24%.
    • Nuclear injection of somatic cell nuclei demonstrated normal developmental patterns of stage-specific epitopes.

    Conclusions:

    • Intracytoplasmic injection of isolated somatic cell nuclei is a relatively efficient method for cloning bovine embryos.
    • The passage number of donor cells did not significantly influence developmental competence.
    • SCNT techniques continue to advance the field of cattle reproductive biotechnology.