Yodo Sugishita1, Naoki Okamoto1, Atsushi Uekawa1, Takayuki Yamochi2, Mariko Nakajima1, Chie Namba1, Suguru Igarashi1, Takumi Sato1, Sei Ohta3, Makoto Takenoshita3, Shu Hashimoto2, Akiko Tozawa4, Yoshiharu Morimoto5, Nao Suzuki6. 1. Department of Obstetrics and Gynecology, St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan. 2. Namba IVF Clinic, Osaka, 550-0015, Japan. 3. Eve Bioscience, Co, Ltd., Wakayama, 648-0003, Japan. 4. Institute of Medical Science, St. Marianna University Graduate School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan. 5. HORAC IVF Grand Front, Osaka, 530-0011, Japan. 6. Department of Obstetrics and Gynecology, St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan. nao@marianna-u.ac.jp.
Abstract
PURPOSE: A device for closed vitrification was designed to reduce the risk of contamination and investigated on its efficacy for ovarian function recovery after cryopreservation and heterotopic transplantation. METHODS: Ovarian tissues from green fluorescence protein transgenic mice (10 GFP mice) were vitrified using the device, and warmed ovarian tissues were transplanted into the ovarian bursa region in wild-type female mice (6 mice). Fresh ovarian tissues were similarly transplanted as a control. After recovery of the estrous cycle, mice were mated with male mice. Ovarian tissues from six cynomolgus monkeys were vitrified and warmed with the device for autologous, heterotopic transplantation. Fresh tissue transplantation was not performed for the control. Ovarian function was examined by recovery of the hormonal cycle. Histological examination was conducted. RESULTS: The number of live pups per recipient mouse was not significantly different after transplantation of fresh or vitrified-warmed ovarian tissue, although the pregnancy rate was reduced with vitrified tissues. The hormonal cycle was restored in 5/6 monkeys after heterotopic transplantation of vitrified-warmed ovarian tissue. Follicles were harvested at eight sites in the omentum and 13 sites in the mesosalpinx. In vitro maturation (IVM)/IVF produced embryo but did not develop. CONCLUSIONS: Resumption of the hormonal cycles, follicle development, and oocyte retrieval from vitrified-warmed ovarian tissue transplants may indicate that the use of vitrification for ovarian tissue in a closed system has a potential of clinical application without the risk of contaminations. More detailed analyses of the effects of vitrification on ovarian tissue, such as gene expression patterns in oocytes and granulosa cells, may be needed for establishing a standard procedure for cryopreservation of ovarian tissues in human.
PURPOSE: A device for closed vitrification was designed to reduce the risk of contamination and investigated on its efficacy for ovarian function recovery after cryopreservation and heterotopic transplantation. METHODS: Ovarian tissues from green fluorescence protein transgenic mice (10 GFP mice) were vitrified using the device, and warmed ovarian tissues were transplanted into the ovarian bursa region in wild-type female mice (6 mice). Fresh ovarian tissues were similarly transplanted as a control. After recovery of the estrous cycle, mice were mated with male mice. Ovarian tissues from six cynomolgus monkeys were vitrified and warmed with the device for autologous, heterotopic transplantation. Fresh tissue transplantation was not performed for the control. Ovarian function was examined by recovery of the hormonal cycle. Histological examination was conducted. RESULTS: The number of live pups per recipient mouse was not significantly different after transplantation of fresh or vitrified-warmed ovarian tissue, although the pregnancy rate was reduced with vitrified tissues. The hormonal cycle was restored in 5/6 monkeys after heterotopic transplantation of vitrified-warmed ovarian tissue. Follicles were harvested at eight sites in the omentum and 13 sites in the mesosalpinx. In vitro maturation (IVM)/IVF produced embryo but did not develop. CONCLUSIONS: Resumption of the hormonal cycles, follicle development, and oocyte retrieval from vitrified-warmed ovarian tissue transplants may indicate that the use of vitrification for ovarian tissue in a closed system has a potential of clinical application without the risk of contaminations. More detailed analyses of the effects of vitrification on ovarian tissue, such as gene expression patterns in oocytes and granulosa cells, may be needed for establishing a standard procedure for cryopreservation of ovarian tissues in human.
Authors: Christiani A Amorim; Mara Curaba; Anne Van Langendonckt; Marie-Madeleine Dolmans; Jacques Donnez Journal: Reprod Biomed Online Date: 2011-05-07 Impact factor: 3.828