title: Freeze-thawing intact human ovary with its vascular pedicle with a passive cooling device.
abstract: OBJECTIVE
  To test the feasibility of freezing intact human ovary using a passive cooling device.
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  Academic research unit of the department of gynecology in a university hospital.
  
  
  METHODS
  Ovaries were obtained from three women who were between 29 and 36 years of age.
  
  
  METHODS
  Ovarian perfusion with cryoprotective solution and slow freezing with a cryofreezing container (5100 Cryo 1 degrees C Freezing Container; Nalgene, VWR, Belgium). Rapid thawing of ovaries by perfusion and bathing with decreased sucrose gradient.
  
  
  METHODS
  Viability of follicles, stromal cells, and vascular components was assessed with live-dead stains in freshly removed ovary, after cryoprotectant exposure before freezing, and after thawing. Histological morphology was assessed at these three different times.
  
  
  RESULTS
  The percentage of live follicles was 99.4% in fresh tissue, 98.1% after cryoprotectant exposure, and 75.1% after thawing. Viability assessment showed live stromal cells and small vessels after thawing. On histological evaluation, the morphology of follicles and cortical and medullar tissue was similar in all three groups.
  
  
  CONCLUSIONS
  We described the cryopreservation, using an accessible protocol, of intact human ovary with its vascular pedicle and proved high survival rates of follicles, small vessels, and stromal cells and a normal histological structure in all the ovarian components after thawing using our protocol.
authors.affiliation: Department of Gynecology, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Belen
authors.initials: B
authors.lastname: Martinez-Madrid
authors.forename: Marie-Madeleine
authors.initials: MM
authors.lastname: Dolmans
authors.forename: Anne
authors.initials: A
authors.lastname: Van Langendonckt
authors.forename: Sylvie
authors.initials: S
authors.lastname: Defrère
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
mesh_terms: D000328:Adult; D001808:Blood Vessels / Q000033:anatomy & histology / Q000166:cytology / Q000502:physiology; D002470:Cell Survival; D015925:Cryopreservation / Q000295:instrumentation*; D004867:Equipment Design; D004864:Equipment and Supplies; D005260:Female; D006801:Humans; D006080:Ovarian Follicle / Q000502:physiology; D010053:Ovary / Q000098:blood supply*; D011446:Prospective Studies; D017154:Stromal Cells / Q000502:physiology; D014022:Tissue Survival