A new step toward the artificial ovary: survival and proliferation of isolated murine follicles after autologous transplantation in a fibrin scaffold

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Presentation transcript:

A new step toward the artificial ovary: survival and proliferation of isolated murine follicles after autologous transplantation in a fibrin scaffold Valérie Luyckx, M.D., Marie-Madeleine Dolmans, M.D., Ph.D., Julie Vanacker, Ph.D., Camille Legat, Cristina Fortuño Moya, B.Sc., Jacques Donnez, M.D., Ph.D., Christiani Andrade Amorim, V.M.D., Ph.D. Fertility and Sterility Volume 101, Issue 4, Pages 1149-1156 (April 2014) DOI: 10.1016/j.fertnstert.2013.12.025 Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Primordial (FP), primary (F1), and secondary (F2) follicles seen after mechanical isolation and (B) their proportion and diameter (mean ± SD). (C) Fibrin clot inside the peritoneal pocket after 7 days of grafting. The clot is easily identified by the nonresorbable suture (faint line around it) used to create the pocket. Fertility and Sterility 2014 101, 1149-1156DOI: (10.1016/j.fertnstert.2013.12.025) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Autografting of murine ovarian follicles in a fibrin clot after 1 week. Some secondary follicles (white arrows) were observed after 1 week of grafting in (A) an F25/T4 fibrin clot and (B) an F12.5/T1 fibrin clot. Antral follicles (black arrows) were evidenced in both formulations: (C) in F25/T4, an ovarian follicle at the beginning of antralization; and (D) in F12.5/T1, ovarian follicles growing up to the antral stage. (E) Proportions of primordial, primary, secondary, and antral follicles and follicular structures found in both fibrin formulations. Fertility and Sterility 2014 101, 1149-1156DOI: (10.1016/j.fertnstert.2013.12.025) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Terminal deoxynucleotide transferase–mediated dUTP nick-end labeling (TUNEL) assay after grafting: 4′,6-diamidino-2-phenylindole counterstaining (cells in blue) and TUNEL staining (cells in red) are merged. (A) Apoptotic granulosa cells stained positive (white arrows) in ovarian follicles and isolated ovarian cells (OCs; white arrowheads); and (B) the grafted fibrin matrix contained only a few apoptotic OCs. (C) Ki67-positive granulosa cells (in brown, white arrows) showing growth of isolated follicles after grafting, and Ki67-positive OCs indicating proliferation of these cells (black arrows). (D) Blood vessels and endothelial cells, stained with anti-CD34, were located around and inside the matrix and surrounding a growing follicle (black arrow). Fertility and Sterility 2014 101, 1149-1156DOI: (10.1016/j.fertnstert.2013.12.025) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 4 CD45-positive cells (in brown) were observed (A) around and infiltrating the fibrin matrix and (B) surrounding a growing follicle. Fertility and Sterility 2014 101, 1149-1156DOI: (10.1016/j.fertnstert.2013.12.025) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions