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Cryobiology2009; 59(1); 36-41; doi: 10.1016/j.cryobiol.2009.04.003

Effect of dehydration prior to cryopreservation of large equine embryos.

Abstract: Cryopreservation of equine embryos>300microm in diameter results in low survival rates using protocols that work well for smaller equine embryos. These experiments tested the potential benefit of incorporating a dehydration step prior to standard cryopreservation procedures. Forty-six, day 7-8, grade 1, equine embryos 300-1350microm in diameter were subjected to one of the following treatments: (A) 2 min in 0.6M galactose, 10min in 1.5M glycerol, slow freeze (n=21); (B) 10min in 1.5M glycerol, slow freeze (n=15); (C) 2min in 0.6M galactose, 10min in 1.5M glycerol, followed by exposure to thaw solutions, then culture medium (n=5); (D) transferred directly to culture medium (n=5). Frozen embryos were thawed and subjected to a three-step cryoprotectant removal. Five embryos from each treatment were evaluated morphologically after 24 and 48h culture (1=excellent, 5=degenerate/dead). All treatments had at least 4/5 embryos with a quality score >or=3 at these time points except treatment B (2/5 at 24h, 1/5 at 48h). Subsequent embryos from treatment A (n=16) or B (n=10) were matched in sets of two for size and treatment, thawed, and immediately transferred in pairs to 13 recipients. Only two recipient mares were pregnant; one received two 400microm embryos from treatment A, and the other one 400 and one 415microm embryo from treatment B. There was no advantage of incorporating a 2min dehydration step into the cryopreservation protocol for large equine embryos.
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Publication Date: 2009-04-16 PubMed ID: 19375416PubMed Central: PMC2706293DOI: 10.1016/j.cryobiol.2009.04.003Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper analyses the effects of a dehydration step before cryopreservation on large equine embryos. The results suggest that inclusion of a 2min dehydration step does not provide any significant benefits to the survival rates of such embryos post cryopreservation.

Methodology and Experimental Groups

  • The researchers worked with 46 grade 1 equine embryos ranging in size from 300-1350 micrometers, all at day 7-8 of development.
  • These embryos were subjected to one of four treatments which varied in terms of exposure to galactose, glycerol, a thaw solution, a slow freeze process, and direct transfer to a culture medium.
  • Embryos were divided and subjected to treatments A (21 embryos), B (15 embryos), C(5 embryos), and D (5 embryos).

Post Treatment Evaluations

  • The embryos were slow frozen and then thawed.
  • The embryos underwent a three-step cryoprotectant removal process.
  • Each group had 5 embryos evaluated morphologically after 24 and 48 hours of culture.
  • The evaluation used a score from 1 (excellent) to 5 (degenerate or dead).
  • All treatments had most of the embryos score at least 3 except for group B which had fewer.

Embryo Transfer

  • Remaining embryos from groups A and B were matched in pairs for size and treatment.
  • These embryos were thawed and immediately transferred into pairs to 13 recipient mares.
  • Out of the 13 recipient mares, only two were pregnant – one that received two 400 micrometer embryos from group A, and the other received one 400 and one 415 micrometer embryo from group B.

Conclusions

  • The researchers concluded that the inclusion of a 2min dehydration step in the cryopreservation protocol for large equine embryos does not confer any significant advantages.
  • Therefore, it seems that such a step might not be necessary for the successful cryopreservation of larger embryos.

Cite This Article

APA
Barfield JP, McCue PM, Squires EL, Seidel GE. (2009). Effect of dehydration prior to cryopreservation of large equine embryos. Cryobiology, 59(1), 36-41. https://doi.org/10.1016/j.cryobiol.2009.04.003

Publication

Cryobiology
ISSN: 1090-2392
NlmUniqueID: 0006252
Country: Netherlands
Language: English
Volume: 59
Issue: 1
Pages: 36-41

Researcher Affiliations

Barfield, J P
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523-1683, USA. jbarfiel@colostate.edu
McCue, P M
    Squires, E L
      Seidel, G E

        MeSH Terms

        • Animals
        • Cryopreservation / methods
        • Cryoprotective Agents / pharmacology
        • Culture Media
        • Embryo Culture Techniques
        • Embryo Transfer / methods
        • Female
        • Fertilization in Vitro / methods
        • Freezing
        • Glycerol / chemistry
        • Horses
        • Pregnancy
        • Pregnancy, Animal
        • Time Factors
        • Water

        Grant Funding

        • T32 HD007031 / NICHD NIH HHS
        • HD07031 / NICHD NIH HHS

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