title: Markers of growth and development in primate primordial follicles are preserved after slow cryopreservation.
abstract: OBJECTIVE
  To investigate the effect of slow cryopreservation on the morphology and function of primate primordial follicles within ovarian tissue slices.
  
  
  METHODS
  Fresh monkey ovarian tissue was frozen by slow cryopreservation and thawed for analysis of morphologic and functional parameters.
  
  
  METHODS
  University-affiliated laboratory.
  
  
  METHODS
  Rhesus monkey ovarian tissue.
  
  
  METHODS
  None.
  
  
  METHODS
  Histologic analysis, follicle counting, assessment of protein abundance and localization.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Center for Reproductive Research, Northwestern University, Evanston, Illinois, USA.
authors.forename: Shiying
authors.initials: S
authors.lastname: Jin
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
authors.forename: Lonnie D
authors.initials: LD
authors.lastname: Shea
authors.forename: Mary B
authors.initials: MB
authors.lastname: Zelinski
authors.forename: Richard L
authors.initials: RL
authors.lastname: Stouffer
authors.forename: Teresa K
authors.initials: TK
authors.lastname: Woodruff
mesh_terms: D028341:Activins / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D049109:Cell Proliferation; D015925:Cryopreservation / Q000379:methods* / Q000662:veterinary; D005260:Female; D008253:Macaca mulatta; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology*; D010053:Ovary / Q000166:cytology*; D010766:Phosphorylation; D021122:Protein Subunits / Q000378:metabolism; D051899:Smad2 Protein / Q000378:metabolism; D046509:Tissue Culture Techniques
chemical_list: D021122:Protein Subunits; D051899:Smad2 Protein; D028341:Activins