title: Chick embryo chorioallantoic membrane (CAM) model: a useful tool to study short-term transplantation of cryopreserved human ovarian tissue.
abstract: OBJECTIVE
  To investigate the chorioallantoic membrane (CAM) model for the study of short-term transplantation of frozen human ovarian tissue.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  Academic research unit.
  
  
  METHODS
  Ovarian tissue was obtained from three women.
  
  
  METHODS
  Frozen-thawed human cortical fragments were grafted onto traumatized CAM or beneath the CAM of 10-day-old chick embryos. Grafts were retrieved after 1, 2, 3, 4, and 5 days in ovo.
  
  
  METHODS
  Viability was assessed by calcein-AM and ethidium homodimer I. Tissue integrity, ischemic injury, and neovascularization were evaluated by histology. Cell proliferation was analyzed by Ki-67 immunohistochemistry.
  
  
  RESULTS
  All the grafts showed adhesion when placed onto CAM, compared with only 30.4% beneath the CAM. Follicles were healthy, apart from a few degenerated follicles in necrotic and fibrotic areas. After 5 days, the majority of follicles were intermediate (32%) or primary (45.7%). Ki-67 immunohistochemistry revealed 12.5% proliferative follicles on day 2, reaching 20.7% on day 5. Fibrosis appeared on day 1; necrosis, follicular degeneration and follicular proliferation on day 2; and neovascularization and stromal cell proliferation on day 3.
  
  
  CONCLUSIONS
  The present study showed that the CAM model provides a new approach to study human ovarian tissue transplantation in its first ischemic stages, yielding information on the timing of tissue changes before the establishment of neovascularization.
authors.affiliation: Department of Gynecology, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Belen
authors.initials: B
authors.lastname: Martinez-Madrid
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Van Eyck
authors.forename: Almudena
authors.initials: A
authors.lastname: Veiga-Lopez
authors.forename: Marie-Madeleine
authors.initials: MM
authors.lastname: Dolmans
authors.forename: Anne
authors.initials: A
authors.lastname: Van Langendonckt
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000502:physiology*; D020256:Choroidal Neovascularization; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D005306:Fertilization / Q000502:physiology; D006801:Humans; D008297:Male; D008954:Models, Biological; D010053:Ovary / Q000098:blood supply / Q000166:cytology* / Q000502:physiology* / Q000637:transplantation