Evaluation of cryopreserved ovarian tissue from prepubertal patients after long-term xenografting and exogenous stimulation Valérie Luyckx, M.D., Sarah.

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Presentation transcript:

Evaluation of cryopreserved ovarian tissue from prepubertal patients after long-term xenografting and exogenous stimulation Valérie Luyckx, M.D., Sarah Scalercio, B.Pharm., M.S., Pascale Jadoul, M.D., Christiani Andrade Amorim, V.M.D., Ph.D., Michelle Soares, M.D., Jacques Donnez, M.D., Ph.D., Marie-Madeleine Dolmans, M.D., Ph.D. Fertility and Sterility Volume 100, Issue 5, Pages 1350-1357.e3 (November 2013) DOI: 10.1016/j.fertnstert.2013.07.202 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Macroscopic view of the surgical procedure. After thawing, two ovarian fragments were grafted intraperitoneally to a female immunodeficient mouse. (B) After long-term grafting and exogenous stimulation, antral follicles were macroscopically observed, measuring 6–8 mm in width. Fertility and Sterility 2013 100, 1350-1357.e3DOI: (10.1016/j.fertnstert.2013.07.202) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Histologic illustrations of ovarian tissue from prepubertal patients before (A and C) and after (B and D) grafting. (A) Histologic illustrations of ovarian tissue from prepubertal patients before grafting. All follicles were inactive and at the primordial stage. Scale bar = 100 μm. (B) After grafting, a few growing follicles were observed (black arrow). However, a large pool of primordial follicles (white arrow) was evidenced in ovarian sections. Scale bar = 100 μm. (C) Histologic illustration of a primordial follicle (white arrow) with a multinucleated oocyte from frozen-thawed prepubertal ovarian tissue. Scale bar = 50 μm. (D) Follicles with multinucleated oocytes and multioocyte follicles were also observed at different follicular stages after grafting. A binucleated oocyte (white arrow) was observed in an antral follicle from prepubertal ovarian tissue after long-term xenografting. Scale bar = 50 μm. Fertility and Sterility 2013 100, 1350-1357.e3DOI: (10.1016/j.fertnstert.2013.07.202) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Graph representing follicular density (expressed as the number of ovarian follicles per mm²) in frozen-thawed and grafted ovarian tissue from prepubertal and adult patients. A significant difference (P<.05) was evidenced between frozen-thawed prepubertal and adult ovarian tissue (A), and also between grafted prepubertal and adult ovarian tissue (B). Fertility and Sterility 2013 100, 1350-1357.e3DOI: (10.1016/j.fertnstert.2013.07.202) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Proportion of primordial and growing (intermediate, primary, secondary, and antral) follicles before and after xenografting in frozen-thawed ovarian fragments from prepubertal patients. a Significantly different (P<.01). b Significantly different (P<.05). Fertility and Sterility 2013 100, 1350-1357.e3DOI: (10.1016/j.fertnstert.2013.07.202) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Histologic illustrations of cryopreserved adult ovarian tissue after thawing (A) and after long-term xenografting (B). (A) A few primordial follicles (white arrows) were evidenced in cryopreserved ovarian tissue after thawing. Scale bar = 100 μm. (B) After transplantation to an immunodeficient mouse, some growing follicles (black arrows) were observed near a few primordial follicles (white arrows). Scale bar = 100 μm. Fertility and Sterility 2013 100, 1350-1357.e3DOI: (10.1016/j.fertnstert.2013.07.202) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 2 Graph showing the proportion of antimüllerian hormone (AMH)-positive follicles in prepubertal ovarian tissue after thawing and after grafting. (2.41% of 2,702 analyzed follicles in frozen-thawed ovarian tissue vs. 16.49% of 624 analyzed follicles in grafted ovarian tissue). Fertility and Sterility 2013 100, 1350-1357.e3DOI: (10.1016/j.fertnstert.2013.07.202) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions