Oocyte retrieval after heterotopic transplantation of ovarian tissue cryopreserved by closed vitrification protocol

封闭式玻璃化冷冻保存卵巢组织异位移植后的卵母细胞提取

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作者:Yodo Sugishita, Naoki Okamoto, Atsushi Uekawa, Takayuki Yamochi, Mariko Nakajima, Chie Namba, Suguru Igarashi, Takumi Sato, Sei Ohta, Makoto Takenoshita, Shu Hashimoto, Akiko Tozawa, Yoshiharu Morimoto, Nao Suzuki

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.

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.

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.

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.

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