Allotransplantation of cryopreserved prepubertal mouse ovaries restored puberty and fertility without affecting methylation profile of Snrpn-DMR Hong-Yan.

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Presentation transcript:

Allotransplantation of cryopreserved prepubertal mouse ovaries restored puberty and fertility without affecting methylation profile of Snrpn-DMR Hong-Yan Wang, M.D., Yun-Hong Li, M.D., Lei Sun, Ph.D., Xuan Gao, M.D., Li You, M.D., Yin Wang, Ph.D., Jing-Long Ma, Ph.D., Zi- Jiang Chen, Ph.D. Fertility and Sterility Volume 99, Issue 1, Pages 241-247.e4 (January 2013) DOI: 10.1016/j.fertnstert.2012.08.030 Copyright © 2013 Terms and Conditions

Figure 1 Viability assay and methylation status of Snrpn imprinted gene of GV oocytes retrieved from fresh and cryopreserved juvenile ovaries. Viable oocytes and methylation rate from (A, D) fresh ovaries, (B, E) vitrified-warmed ovaries with ED20 solution, and (C, F) vitrified-warmed ovaries with EG5.5/30 solution. Red fluorescence of dead cells and green fluorescence of live cells. Bar = 50 μm. Fertility and Sterility 2013 99, 241-247.e4DOI: (10.1016/j.fertnstert.2012.08.030) Copyright © 2013 Terms and Conditions

Figure 2 Metaphase II oocytes originating from fresh and cryopreserved transplanted ovaries further developed to blastocysts. Shown are the mature oocytes and early embryos derived from fresh ovaries (A–D), vitrified-warmed ovaries treated with ED20 solution (E–H), and vitrified-warmed ovaries treated with EG5.5/30 solution (I–L). Metaphase I and II oocytes were collected from antral follicles of fresh or vitrified-warmed surviving ovaries by superovulation; only metaphase II oocytes underwent IVF and early embryonic development (A, E, I). There were no remarkable morphological differences among the three groups. Arrowheads (A, E, I) show first polar bodies in metaphase II oocytes. Bar = 100 μm. Fertility and Sterility 2013 99, 241-247.e4DOI: (10.1016/j.fertnstert.2012.08.030) Copyright © 2013 Terms and Conditions

Supplemental Figure 1 Histological analysis of fresh and cryopreserved juvenile mouse ovaries. Histology observation in (A) fresh, (B) vitrification with ED20 solution, and (C) vitrification with EG5.5/30 solution. Primordial and growing follicles from vitrified-warmed ovaries showed good morphology compared with those from fresh ones (black arrow: primordial follicle; red arrow: growing follicle). Bar = 50 μm. Fertility and Sterility 2013 99, 241-247.e4DOI: (10.1016/j.fertnstert.2012.08.030) Copyright © 2013 Terms and Conditions

Supplemental Figure 2 Histological and macroscopic appearance of surviving ovarian grafts in the fresh and cryopreserved transplanted groups. (A) The vitrified-warmed juvenile ovary was laid beneath the renal capsule of the recipient mouse (arrow); (B, C) the surviving ovarian grafts were evidently increased under renal capsule (B: red arrow: follicles; black arrow: blood vessel; C: red arrow: corpus rubrum); (D) histological appearance of the fresh graft and (E) histological appearance of the vitrified graft treated with ED20 solution; (F) 2 months after the transplantation, macroscopic appearance of the surviving graft treated with EG5.5/30 solution under renal capsule (arrow: follicles); (A and B, C, F, bar = 10 mm; C and D, bar = 100 μm) CL = corpus luteum; Ant.F = antral follicle; Sec.F = secondary follicle. Fertility and Sterility 2013 99, 241-247.e4DOI: (10.1016/j.fertnstert.2012.08.030) Copyright © 2013 Terms and Conditions