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Related Experiment Videos

Blastocyst viability and generation of transgenic cattle following freezing of in vitro produced, DNA-injected

Y M Han1, S J Kim, J S Park

  • 1Korea Research Institute of Bioscience and Biotechnology, P.O. Box 115, Yusong, 305-600, Taejon, South Korea.

Animal Reproduction Science
|September 1, 2000
PubMed
Summary

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Freezing impacts DNA-injected bovine embryo viability, with quality and developmental stage influencing survival rates. However, frozen embryos can still produce transgenic calves, demonstrating feasibility for genetic applications.

Area of Science:

  • Reproductive Biology
  • Embryology
  • Genetics

Background:

  • In vitro production of bovine embryos is crucial for genetic applications.
  • Cryopreservation is essential for long-term storage and transport of embryos.
  • Assessing the viability of frozen, genetically modified embryos is critical for successful application.

Purpose of the Study:

  • To determine the effect of cryopreservation on the in vitro viability of DNA-injected bovine embryos.
  • To evaluate the in vivo developmental potential of frozen-thawed DNA-injected bovine embryos.
  • To assess the efficiency of generating transgenic cattle from frozen embryos.

Main Methods:

  • In vitro fertilization and DNA injection (pBL1 gene) of bovine zygotes.
  • Co-culture with mouse embryonic fibroblasts (MEF) in CR1aa medium.

Related Experiment Videos

  • Conventional slow freezing and rapid thawing protocols for cryopreservation.
  • In vitro assessment of embryo survival and morphological quality.
  • Embryo transfer to recipient cows to determine in vivo viability and pregnancy rates.
  • Analysis of transgene transmission in offspring.

    • Main Results:

    • Excellent quality DNA-injected blastocysts exhibited higher post-thaw survival rates (78.8%) compared to good (60.9%) or fair (12.5%) quality embryos.
    • Blastocyst survival increased with developmental stage, with expanded blastocysts showing 71.2% survival.
    • Pregnancy rates were lower for frozen embryos (13.6%) than fresh embryos (26.5%).
    • Two transgenic male calves were born from frozen-thawed embryos, with one demonstrating germline mosaicism (13.0% transmission rate).

    Conclusions:

    • Cryopreservation significantly affects the viability of DNA-injected bovine embryos, with quality and developmental stage being critical factors.
    • Despite reduced pregnancy rates, frozen-thawed embryos can successfully produce transgenic cattle.
    • This study confirms the feasibility of cryopreserving DNA-injected, in vitro produced bovine embryos for generating genetically modified livestock.