Chick embryo chorioallantoic membrane (CAM) model: a useful tool to study short-term transplantation of cryopreserved human ovarian tissue.
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.
Martinez-Madrid
Belen
B
Department of Gynecology, Université Catholique de Louvain, Brussels, Belgium.
Donnez
Jacques
J
Van Eyck
Anne-Sophie
AS
Veiga-Lopez
Almudena
A
Dolmans
Marie-Madeleine
MM
Van Langendonckt
Anne
A
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