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Related Experiment Video

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Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification
06:00

Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification

Published on: November 18, 2011

Equilibrium vitrification of mouse embryos.

Bo Jin1, Keiji Mochida, Atsuo Ogura

  • 1Laboratory of Animal Science, College of Agriculture, Kochi University, Nankoku, Kochi, Japan.

Biology of Reproduction
|October 16, 2009
PubMed
Summary

A novel cryopreservation method achieves near-equilibrium vitrification for embryos, improving survival rates after prolonged storage at -80°C and even after rewarming from liquid nitrogen.

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Area of Science:

  • Reproductive Biology
  • Cryobiology
  • Developmental Biology

Background:

  • Vitrification is a common embryo cryopreservation technique with advantages but suffers from nonequilibrium cooling.
  • Supercooling during vitrification can lead to intracellular ice formation and reduced embryo survival.

Purpose of the Study:

  • To develop a novel embryo cryopreservation method using near-equilibrium vitrification.
  • To assess embryo survival after near-equilibrium vitrification and prolonged cold storage.

Main Methods:

  • Two-cell ICR mouse embryos were vitrified using ethylene glycol (EG)-based solutions (EFSa, EFSc) with varying EG and sucrose concentrations.
  • Embryo survival was evaluated after rapid warming, storage at -80°C for 4 or 10 days, and after recooling in liquid nitrogen.

Main Results:

  • High in vitro survival rates were observed for embryos vitrified with 30%-40% EG solutions upon rapid warming.
  • Significant embryo survival was achieved after storage at -80°C for 4 and 10 days using specific EG-sucrose solutions (EFS30c, EFS40c, EFS35c).
  • Vitrified embryos (ICR, C57BL/6J) maintained high survival after liquid nitrogen recooling, with 75% of C57BL/6J embryos developing to term.

Conclusions:

  • A novel near-equilibrium vitrification method for embryo cryopreservation was successfully developed.
  • This technique enhances embryo survival after extended cold storage and extreme temperature fluctuations.
  • The method combines the benefits of vitrification and equilibrium slow freezing for superior cryopreservation outcomes.