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Related Experiment Videos

Morphological changes in mouse embryos cryopreserved by different techniques.

A R S Coutinho1, C M Mendes, H V A Caetano

  • 1Department of Animal Reproduction, School of Veterinary Medicine, University of São Paulo, São Paulo, Brazil. manicout@usp.br

Microscopy Research and Technique
|December 21, 2006
PubMed
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Vitrification causes more severe cellular damage and reduces viability in cryopreserved mammalian embryos compared to slow freezing. Both methods induce apoptosis and oncosis, but vitrification

Area of Science:

  • Reproductive Biotechnology
  • Cell Biology
  • Histology

Background:

  • Cryopreservation is vital for mammalian embryo applications.
  • Assessing embryo viability often relies on subjective methods.
  • Understanding cryopreservation-induced cellular injury is crucial for optimizing protocols.

Purpose of the Study:

  • To compare morphological and ultrastructural alterations in mouse embryos after slow freezing versus vitrification.
  • To evaluate embryo viability using membrane integrity assays and microscopy techniques.

Main Methods:

  • Comparison of fresh, slow-frozen, and vitrified mouse embryos.
  • Assessment of cell membrane integrity using Hoechst33342/propidium iodide (H/PI) staining.
  • Morphological analysis via hematoxylin and eosin (HE) staining on historesin-embedded samples.

Related Experiment Videos

  • Ultrastructural evaluation using transmission electron microscopy.

    • Main Results:

    • Vitrification resulted in significantly higher membrane permeability (69.8%) than slow freezing (48.4%) or controls (13.8%).
    • Historesin embedding facilitated superior morphological assessment, revealing nuclear pycnosis in both cryopreservation groups.
    • Slow freezing induced apoptosis (cytoplasmic condensation, eosinophilia), while vitrification led to oncosis (degenerated cells).

    Conclusions:

    • Both slow freezing and vitrification induce apoptosis and oncosis in mammalian embryos.
    • Vitrification causes more significant cellular damage and reduced viability compared to slow freezing.
    • Historesin embedding is recommended for detailed morphological analysis of cryopreserved embryos.