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Related Experiment Videos

New preservation method for mouse spermatozoa without freezing.

Nguyen Van Thuan1, Sayaka Wakayama, Satoshi Kishigami

  • 1RIKEN Kobe Institute, Center for Developmental Biology, Laboratory for Genomic Reprogramming, Chuo-ku, Kobe City, Hyogo 650-0047, Japan. nvthuan@cdb.riken.jp

Biology of Reproduction
|October 16, 2004
PubMed
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This summary is machine-generated.

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Mouse sperm can be preserved at 4 degrees C in a high-osmolarity medium for extended periods, enabling successful fertilization and live births via intracytoplasmic sperm injection (ICSI). This method offers a viable alternative to freezing for sperm preservation.

Area of Science:

  • Reproductive Biology
  • Sperm Cryopreservation Alternatives
  • Mammalian Embryology

Background:

  • Traditional sperm preservation often involves freezing, which can damage sperm.
  • Developing non-freezing methods is crucial for maintaining sperm viability and function.
  • Intracytoplasmic sperm injection (ICSI) requires high-quality, viable sperm for successful fertilization.

Purpose of the Study:

  • To investigate non-freezing preservation methods for mouse spermatozoa.
  • To evaluate the impact of preserved sperm on oocyte fertilization and subsequent embryonic development.
  • To identify optimal conditions for sperm storage to maximize live birth rates after ICSI.

Main Methods:

  • Sperm were stored in KSOMaa medium with varying BSA concentrations and temperatures (4°C, RT, 37°C).

Related Experiment Videos

  • High osmolarity (271-2000 mOsmol) KSOM-BSA media at 4°C were tested.
  • A two-step preservation protocol (RT then -20°C) was evaluated.
  • Preserved sperm were used for ICSI in mature mouse oocytes.

    • Main Results:

    • Preservation at 4°C in high osmolarity (700-1000 mOsmol) KSOM-BSA yielded the highest live birth rates.
    • Optimal conditions identified: 800 mOsmol KSOM-BSA at 4°C.
    • Sperm stored for 2 months under optimal conditions resulted in >40% morula/blastocyst development and 39% live births.
    • A two-step method allowed storage for 3 months at -20°C after 1 week at RT, retaining ICSI competence.

    Conclusions:

    • Non-freezing preservation of mouse spermatozoa is feasible and effective.
    • High osmolarity (800 mOsmol) and 4°C storage in KSOM-BSA are optimal for sperm preservation.
    • These methods support normal embryonic development and healthy offspring production via ICSI.
    • Extended sperm conservation without freezing is achievable, offering a valuable alternative for reproductive technologies.