Pup birth from mouse oocytes in preantral follicles derived from vitrified and warmed ovaries followed by in vitro growth, in vitro maturation, and in.

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Pup birth from mouse oocytes in preantral follicles derived from vitrified and warmed ovaries followed by in vitro growth, in vitro maturation, and in vitro fertilization Akiko Hasegawa, Ph.D., Nahoko Mochida, M.Sc., Toshitada Ogasawara, M.D., Ph.D., Koji Koyama, M.D., Ph.D. Fertility and Sterility Volume 86, Issue 4, Pages 1182-1192 (October 2006) DOI: 10.1016/j.fertnstert.2005.12.082 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Influences of cryoprotectant toxicity and vitrification–warming on mouse ovaries on histological examination. (A) Paraffin section from fresh mouse ovary. (B) Ovarian section treated with equilibration and vitrification solutions followed by thawing process without immersion in liquid nitrogen. (C) Ovarian section treated with a standard method of vitrification and warming as shown in the Materials and Methods section. The three mouse ovarian sections are shown to be of a comparatively similar morphological appearance. Bar = 100 μm. Hasegawa. Pup birth from cryopreserved ovaries. Fertil Steril 2006. Fertility and Sterility 2006 86, 1182-1192DOI: (10.1016/j.fertnstert.2005.12.082) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Histological analysis of vitrified–warmed ovaries in rabbits. (A, C) Fresh ovarian sections. (B, D) Vitrified–warmed sections. Primordial and growing follicles from vitrified–warmed ovaries showed good morphology compared with those from fresh ones. No significant differences were observed in primordial and antral follicles between fresh and vitrified–warmed ovaries (primordial follicle: arrowhead; growing follicle: black arrow; antral follicle: red arrow). The follicles at various stages preserved good status. Bar = 100 μm. Hasegawa. Pup birth from cryopreserved ovaries. Fertil Steril 2006. Fertility and Sterility 2006 86, 1182-1192DOI: (10.1016/j.fertnstert.2005.12.082) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Histological analysis of pig ovarian sections after cryopreservation. (A) Fresh ovarian section. (B, C) Vitrified–warmed ovarian sections. Primordial and growing follicles (B, C) preserved a normal appearance compared with the fresh ovarian section (A). Oocytes and the surrounding granulosa cells showed good morphology, although disconnection (arrows) was observed in some growing follicles after cryopreservation (C). Bar = 100 μm. Hasegawa. Pup birth from cryopreserved ovaries. Fertil Steril 2006. Fertility and Sterility 2006 86, 1182-1192DOI: (10.1016/j.fertnstert.2005.12.082) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Mature oocytes and further developed embryos originating from preantral follicles of fresh and vitrified–warmed ovaries. (A–D) The mature oocytes and embryos derived from the vitrified–warmed ovaries. (E–H) The mature oocytes and embryos of the fresh ovaries. Oocyte–granulosa cell complexes were collected from preantral follicles in fresh or vitrified–warmed ovaries and subjected to IVG-IVM, IVF, and further cultivation. There were not morphologically remarkable differences between the two groups. Arrowheads (A, E) show first polar bodies in metaphase II oocytes. Hasegawa. Pup birth from cryopreserved ovaries. Fertil Steril 2006. Fertility and Sterility 2006 86, 1182-1192DOI: (10.1016/j.fertnstert.2005.12.082) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 5 Live pups delivered by cesarean section (A) and implanted uterus (B). Red arrows, placenta for the live pups. In addition, six traces of implantation were observed (arrows). Hasegawa. Pup birth from cryopreserved ovaries. Fertil Steril 2006. Fertility and Sterility 2006 86, 1182-1192DOI: (10.1016/j.fertnstert.2005.12.082) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions