title: Vitrification and xenografting of human ovarian tissue.
abstract: OBJECTIVE
  To assess the efficiency of two vitrification protocols to cryopreserve human preantral follicles with the use of a xenografting model.
  
  
  METHODS
  Pilot study.
  
  
  METHODS
  Gynecology research unit in a university hospital.
  
  
  METHODS
  Ovarian biopsies were obtained from seven women aged 30-41 years.
  
  
  METHODS
  Ovarian tissue fragments were subjected to one of three cryopreservation protocols (slow freezing, vitrification protocol 1, and vitrification protocol 2) and xenografted for 1 week to nude mice.
  
  
  METHODS
  The number of morphologically normal follicles after cryopreservation and grafting and fibrotic surface area were determined by histologic analysis. Apoptosis was assessed by the TUNEL method. Morphometric analysis of TUNEL-positive surface area also was performed. Follicle proliferation was evaluated by immunohistochemistry.
  
  
  RESULTS
  After xenografting, a difference was observed between the cryopreservation procedures applied. According to TUNEL analysis, both vitrification protocols showed better preservation of preantral follicles than the conventional freezing method. Moreover, histologic evaluation showed a significantly higher proportion of primordial follicles in vitrified (protocol 2)-warmed ovarian tissue than in frozen-thawed tissue. The proportion of growing follicles and fibrotic surface area was similar in all groups.
  
  
  CONCLUSIONS
  Vitrification procedures appeared to preserve not only the morphology and survival of preantral follicles after 1 week of xenografting, but also their ability to resume folliculogenesis. In addition, vitrification protocol 2 had a positive impact on the quiescent state of primordial follicles after xenografting.
authors.affiliation: Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Christiani Andrade
authors.initials: CA
authors.lastname: Amorim
authors.forename: Marie-Madeleine
authors.initials: MM
authors.lastname: Dolmans
authors.forename: Anu
authors.initials: A
authors.lastname: David
authors.forename: Jonathan
authors.initials: J
authors.lastname: Jaeger
authors.forename: Julie
authors.initials: J
authors.lastname: Vanacker
authors.forename: Alessandra
authors.initials: A
authors.lastname: Camboni
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
authors.forename: Anne
authors.initials: A
authors.lastname: Van Langendonckt
mesh_terms: D000328:Adult; D000818:Animals; D017209:Apoptosis; D001706:Biopsy; D049109:Cell Proliferation; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D053960:DNA Breaks; D005260:Female; D005355:Fibrosis; D006085:Graft Survival; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D051379:Mice; D008819:Mice, Nude; D006080:Ovarian Follicle / Q000187:drug effects / Q000473:pathology / Q000637:transplantation*; D010053:Ovary / Q000187:drug effects / Q000473:pathology / Q000637:transplantation*; D010865:Pilot Projects; D017154:Stromal Cells / Q000473:pathology / Q000637:transplantation; D013997:Time Factors; D014183:Transplantation, Heterologous; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents