Markers of growth and development in primate primordial follicles are preserved after slow cryopreservation

Fertil Steril. 2010 May 15;93(8):2627-32. doi: 10.1016/j.fertnstert.2009.11.029. Epub 2010 Jan 13.

Abstract

Objective: To investigate the effect of slow cryopreservation on the morphology and function of primate primordial follicles within ovarian tissue slices.

Design: Fresh monkey ovarian tissue was frozen by slow cryopreservation and thawed for analysis of morphologic and functional parameters.

Setting: University-affiliated laboratory.

Animals: Rhesus monkey ovarian tissue.

Intervention(s): None.

Main outcome measure(s): Histologic analysis, follicle counting, assessment of protein abundance and localization.

Result(s): After freezing and thawing, 89% of the primordial follicles maintained their laminar-based architecture, with sizes close to those of fresh fixed follicles. Molecular markers of early follicle health (activin subunits and the phosphorylated form of the signaling protein Smad2 [pSmad2]) were present in fresh and frozen-thawed primordial follicles. Stroma cells, but not follicles, had a higher level of TUNEL staining. Granulosa cells within the follicles of frozen-thawed ovarian tissue cultured for 48 hours had the capacity to proliferate and sustained expression of the activin subunits and nuclear pSmad2.

Conclusion(s): This study provides evidence that markers of early follicle growth and development are preserved after slow cryopreservation and thaw, with little effect on follicle morphology and function.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Activins / metabolism
  • Animals
  • Apoptosis
  • Cell Proliferation
  • Cryopreservation / methods*
  • Cryopreservation / veterinary
  • Female
  • Macaca mulatta
  • Ovarian Follicle / cytology
  • Ovarian Follicle / physiology*
  • Ovary / cytology*
  • Phosphorylation
  • Protein Subunits / metabolism
  • Smad2 Protein / metabolism
  • Tissue Culture Techniques

Substances

  • Protein Subunits
  • Smad2 Protein
  • Activins