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A protocol used for splitting mouse embryos into two halves.

K P Agrawal, C Polge

    Indian Journal of Experimental Biology
    |July 1, 1989
    PubMed
    Summary
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    Researchers successfully split mouse embryos at the 8-16 cell stage and early blastocyst stage. While splitting was easier in 8-16 cell embryos, both stages showed subsequent development in culture.

    Area of Science:

    • Developmental Biology
    • Embryology
    • Reproductive Science

    Background:

    • Embryo splitting is a technique with potential applications in animal breeding and research.
    • Understanding the optimal stage for embryo manipulation is crucial for successful assisted reproductive technologies.

    Purpose of the Study:

    • To investigate the feasibility and success rate of splitting mouse embryos at different developmental stages (8-16 cell vs. early blastocyst).
    • To assess the subsequent developmental potential of bisected mouse embryos cultured in vitro.

    Main Methods:

    • Obtained 8-16 cell embryos and early blastocysts from albino mice 70 and 90 hours post-LH injection.
    • Utilized a micromanipulator with microtools for embryo bisection.
    • Cultured bisected embryos in T-6 medium within a CO2 incubator (39°C, 5% CO2).

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    Main Results:

    • Splitting was more successful in 8-16 cell embryos (52.44%) compared to early blastocysts (35.48%).
    • Following bisection, 38.88% of 8-16 cell embryos developed further in culture over 48 hours.
    • Only 11.36% of bisected early blastocysts demonstrated development after 48 hours of culture.

    Conclusions:

    • The 8-16 cell stage appears more amenable to successful embryo splitting and subsequent in vitro development in mice.
    • This technique holds promise for applications in animal embryo research and commercial breeding programs.