Successful vitrification of bovine blastocysts, derived by in vitro maturation and fertilization

Molecular Reproduction and Development - Tập 34 Số 3 - Trang 266-271 - 1993
S. Tachikawa1, Takeshige Otoi1, Sosuke Kondo1, T. Machida2, Magosaburo Kasai2
1Tokushima Prefectural Beef and Swine Experiment Station, Anan, Tokushima
2Laboratory of Animal Science, College of Agriculture, Kochi University, Nankoku, Kochi, Japan

Tóm tắt

AbstractBovine blastocysts were produced through maturation, fertilization, and development in vitro. For vitrification, solutions designated EFS, GFS, and PFS were prepared; these were 40% ethylene glycol, 40% glycerol, and 40% propylene glycol, respectively, diluted in modified phosphate‐buffered saline (PBS) containing 30% Ficoll + 0.5 M sucrose. The embryos were exposed to the solutions in one step at room temperature, kept in the solutions for various times, vitrified in liquid nitrogen, and warmed rapidly. When the embryos were vitrified in EFS solution after 1 or 2 min exposure, the postwarming survival rate, assessed by the reexpansion of the blastocoel, was 74–77%. However, when the exposure time was extended to 3 min or longer, this rate dropped to 7–0%. This reduction was attributed to the toxicity of ethylene glycol. Of the embryos vitrified in GFS solution, 53% survived when they were cooled after 1 min exposure; as the duration of the exposure increased, the survival rate increased, reaching a peak (72%) at 4 min. The rate then decreased gradually with exposure time. In PFS solution, embryos surviving after vitrification were recovered only with 1 min exposure (33%), reflecting the high toxicity of propylene glycol. After vitrification in EFS or GFS solution, two embryos were nonsurgically transferred into each of 14 recipient animals. Of the 14 recipients, ten (71%) became pregnant; two resulted in early stillbirths, four recipients delivered twins (four alive and four stillborn), and two delivered single live calves, demonstrating the effectiveness of this simple vitrification method for the cryopreservation of in‐vitro‐produced bovine blastocysts. © 1993 Wiley‐Liss, Inc.

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Molecular Reproduction and Development

Tập 34 Số 3

266-271

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