A new step toward the artificial ovary: survival and proliferation of isolated murine follicles after autologous transplantation in a fibrin scaffold.
OBJECTIVE
To create an artificial ovary to provide an alternative way of restoring fertility in patients who cannot benefit from transplantation of cryopreserved ovarian tissue due to the threat of reintroducing malignant cells.


METHODS
In vivo experimental study.


METHODS
Gynecology research unit in a university hospital.


METHODS
Six-week-old female NMRI mice.


METHODS
Autografting of isolated preantral follicles and ovarian cells (OCs) encapsulated in two fibrin matrices containing low concentrations of fibrinogen (F; mg/mL) and thrombin (T; IU/mL): F12.5/T1 and F25/T4.


METHODS
Follicular density and development, OC survival and proliferation, inflammatory response, and vascularization.


RESULTS
After 1 week, the follicle recovery rate ranged from 30.8% (F25/T4) to 31.8% (F12.5/T1). With both fibrin formulations, all follicles were found to be alive or minimally damaged, as demonstrated by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay, and at the growing stage (primary, secondary, and antral follicles), confirmed by Ki67 immunostaining. Isolated OCs also survived and proliferated after grafting, as evidenced by <1% apoptotic cells and a high proportion of Ki67-positive cells. Vessels were found in both fibrin formulations, and the global vascular surface area varied from 1.35% (F25/T4) to 1.88% (F12.5/T1). Numerous CD45-positive cells were also observed in both F25/T4 and F12.5/T1 combinations.


CONCLUSIONS
The present study is the first to show survival and growth of isolated murine ovarian follicles 1 week after autotransplantation of isolated OCs in a fibrin scaffold. The results indicate that fibrin is a promising candidate as a matrix for the construction of an artificial ovary. Xenotransplantation of isolated human follicles and OCs is the necessary next step to validate these findings.
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Valérie
V
Luyckx
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium; Gynecology Department, Cliniques Universitaires Saint-Luc, Brussels, Belgium. Electronic address: marie-madeleine.dolmans@uclouvain.be.
Marie-Madeleine
MM
Dolmans
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Julie
J
Vanacker
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Camille
C
Legat
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Cristina
C
Fortuño Moya
Society for Research into Infertility, Brussels, Belgium.
Jacques
J
Donnez
Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Christiani Andrade
CA
Amorim
D000818:Animals; D021522:Bioartificial Organs; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005260:Female; D005337:Fibrin / Q000737:chemistry*; D051379:Mice; D006080:Ovarian Follicle / Q000166:cytology / Q000254:growth & development* / Q000637:transplantation*; D010053:Ovary / Q000166:cytology* / Q000254:growth & development*; D054457:Tissue Scaffolds
D005337:Fibrin
Preantral follicles; artificial ovary; cancer patients; fertility preservation; fibrin; ovarian cells; transplantation