Immunohistochemical localization of growth factors after cryopreservation and 3 weeks' xenotransplantation of human ovarian tissue Anu David, M.Sc.,

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Immunohistochemical localization of growth factors after cryopreservation and 3 weeks' xenotransplantation of human ovarian tissue Anu David, M.Sc., Marie-Madeleine Dolmans, M.D., Ph.D., Anne Van Langendonckt, Ph.D., Jacques Donnez, M.D., Ph.D., Christiani Andrade Amorim, V.M.D., Ph.D. Fertility and Sterility Volume 95, Issue 4, Pages 1241-1246 (March 2011) DOI: 10.1016/j.fertnstert.2010.06.007 Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Follicular proportion expressed as a percentage before and after grafting of human ovarian tissue. Although the proportion of growing follicles statistically significantly increased after xenografting, there was no statistically significant difference with respect to primordial and growing follicles between fresh and frozen-thawed grafts. Primordial follicles, fresh vs. fresh graft: P<.05; frozen vs. frozen graft: P<.01. Growing follicles, fresh vs. fresh graft: P<.05; frozen vs. frozen graft: P<.01. Fertility and Sterility 2011 95, 1241-1246DOI: (10.1016/j.fertnstert.2010.06.007) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Immunohistochemical staining for c-kit. (A) Oolemma of primordial and primary follicles in fresh ovarian tissue (23 μm). (B) Faint staining of primordial and primary follicles in fresh grafts (26 μm). (C) Oolemma of primordial follicles in frozen-thawed grafts (19 μm). (D) Negative control (25 μm). (E) Positive control. Fertility and Sterility 2011 95, 1241-1246DOI: (10.1016/j.fertnstert.2010.06.007) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Immunohistochemical staining for dispersed kit ligand (KL) in oocytes and granulosa cells. (A) Primary follicle in fresh ovarian tissue (27 μm). (B) Primary follicle in fresh grafts (24 μm). (C) Transitional and primary follicles in frozen-thawed grafts (31 μm). (D) Negative control (23 μm). (E) Positive control. Fertility and Sterility 2011 95, 1241-1246DOI: (10.1016/j.fertnstert.2010.06.007) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Immunohistochemistry staining for growth differentiation factor-9 (GDF-9). (A) Oocyte from a primary follicle in fresh ovarian tissue (34 μm). (B) Oocyte from a primary follicle in a fresh graft (29 μm). (C) Oocyte from a primary follicle in frozen-thawed graft (28 μm). (D) Negative control (38 μm). (E) Positive control. Fertility and Sterility 2011 95, 1241-1246DOI: (10.1016/j.fertnstert.2010.06.007) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions