Qualitative and morphometric analysis of the ultrastructure of human oocytes cryopreserved by two alternative slow cooling protocols

J Assist Reprod Genet. 2010 Apr;27(4):131-40. doi: 10.1007/s10815-010-9394-7. Epub 2010 Feb 23.

Abstract

Purpose: To ascertain possible cell damage from cryopreservation, the ultrastructure of human oocytes cryopreserved by slow cooling was assessed.

Materials and methods: Cryopreservation was performed through two protocols with one-step or two-step propanediol. Fresh control oocytes were examined for comparison. Samples were processed for transmission electron microscopy analysis.

Results: By light microscopy, both fresh and frozen-thawed oocytes appeared regularly rounded, with intact zona pellucida, and homogeneous cytoplasm. By electron microscopy observation, organelles were abundant and uniformly dispersed. Mitochondria-smooth endoplasmic reticulum associations appeared regular. However, both the amount and density of cortical granules appeared abnormally reduced in frozen-thawed samples. Slight to moderate vacuolization was also found in the ooplasm of oocytes of both frozen groups.

Conclusions: Slow cooling ensures a good overall preservation of human oocytes. However, cytoplasmic vacuolization and cortical granule loss appears associated with cryopreservation, irrespective of the protocol used.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cryopreservation*
  • Cryoprotective Agents / pharmacology*
  • Cytoplasmic Granules / ultrastructure
  • Endoplasmic Reticulum, Smooth / ultrastructure
  • Female
  • Freezing
  • Humans
  • Microscopy, Electron, Transmission
  • Mitochondria / ultrastructure
  • Oocytes*
  • Organ Preservation*
  • Propylene Glycols / pharmacology*

Substances

  • Cryoprotective Agents
  • Propylene Glycols