title: Cryopreservation and xenotransplantation of human ovarian tissue: an ultrastructural study.
abstract: OBJECTIVE
  To analyze the ultrastructure of human ovarian follicles after cryopreservation and short-term xenografting.
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  Academic gynecology and anatomy research units.
  
  
  METHODS
  Ovarian cortical biopsy specimens were obtained from 13 patients.
  
  
  METHODS
  Each ovarian biopsy specimen was dissected into pieces of 1 mm(3) and divided into three groups: [1] fresh tissue, [2] frozen-thawed tissue, and [3] frozen-thawed tissue xenografted onto the peritoneum of nude mice for 3 weeks.
  
  
  METHODS
  Follicular ultrastructure was assessed by light and transmission electron microscopy in [1] fresh, [2] frozen, and [3] frozen-grafted tissue.
  
  
  RESULTS
  Thirty-five ovarian follicles were analyzed by light and transmission electron microscopy. Twenty-five primordial and primary ovarian follicles were found. Most of them exhibited ultrastructurally well preserved features (fresh [N = 8/10], frozen [N = 7/10], and frozen-grafted [N = 4/5] tissue). Ten secondary follicles were present in xenografts. By transmission electron microscopy, all the healthy-looking secondary follicles (70%) were shown to contain intact oocytes, with features typical of earlier developmental stages, surrounded by several layers of follicular cells.
  
  
  CONCLUSIONS
  The present study demonstrates, for the first time, that cryopreservation and xenotransplantation do not appear to greatly affect human primordial/primary follicle ultrastructure. Interestingly, in frozen-thawed xenografts, secondary human ovarian follicles presented a well preserved ultrastructure, but asynchrony between oocyte and granulosa cell development was detected. The possible causes for this asynchrony are discussed.
authors.affiliation: Department of Anatomy, University of Rome "La Sapienza," Rome, Italy.
authors.forename: Stefania A
authors.initials: SA
authors.lastname: Nottola
authors.forename: Alessandra
authors.initials: A
authors.lastname: Camboni
authors.forename: Anne
authors.initials: A
authors.lastname: Van Langendonckt
authors.forename: Dominique
authors.initials: D
authors.lastname: Demylle
authors.forename: Guido
authors.initials: G
authors.lastname: Macchiarelli
authors.forename: Marie-Madeleine
authors.initials: MM
authors.lastname: Dolmans
authors.forename: Belen
authors.initials: B
authors.lastname: Martinez-Madrid
authors.forename: Silvia
authors.initials: S
authors.lastname: Correr
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
mesh_terms: D000328:Adult; D000818:Animals; D015925:Cryopreservation; D005260:Female; D006107:Granulosa Cells / Q000637:transplantation / Q000648:ultrastructure; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D046529:Microscopy, Electron, Transmission; D009865:Oocytes / Q000637:transplantation / Q000648:ultrastructure; D006080:Ovarian Follicle / Q000637:transplantation* / Q000648:ultrastructure*; D010537:Peritoneum / Q000601:surgery; D013997:Time Factors; D014183:Transplantation, Heterologous