Nobuya Aono

Cryopreservation and in vitro maturation of germinal vesicle stage oocytes of animals for application in assisted reproductive technology

Cryopreservation, in vitro maturation, fertilization and culture can be applied to various processes across a wide range of species, that is, for the breeding and reproduction of farm animals, preservation of genetic variants in laboratory animals, and the conservation of wild species. In particular, the storage of oocytes by cryopreservation and IVM following cryopreservation, might become effective alternative assisted reproduction treatments for infertile patients. For example, in a clinical context, these techniques might be important for patients who are at risk of losing their ovarian function because of extirpative therapy, chemotherapy or radiation. Thus, it is important for assisted reproductive technology to improve IVM and cryopreservation techniques. In the present review, we introduce our recent studies on vitrification and IVM of germinal vesicle stage oocytes in animals.

Residual ethylene glycol and dimethyl sulphoxide concentration in human ovarian tissue during warming/thawing steps following cryopreservation

There have been 60 births after transplantation of cryopreserved ovarian tissue: 58 using the slow freezing method, and two using the vitrification method. DMSO and EG are widely used as cryoprotectants. However DMSO is a known epimutagen, and EG has been reported to be toxic in high concentrations. In this study, we measured residual DMSO and EG in ovarian tissue after vitrification and slow freezing. Cryoprotectants remained at a high concentration in the vitrified/warmed ovarian tissue just before transplantation (DMSO: 9.8 mg/g, EG: 9.8 mg/g). We must consider the impact of the cryoprotectants on the mother and the baby.

Efficacy of the endometrial receptivity array for repeated implantation failure in Japan: A retrospective, two-centers study

This study aimed to assess the efficacy of the endometrial receptivity array (ERA) as a diagnostic tool and the impact of personalized embryo transfer (pET) for the treatment of patients with recurrent implantation failure (RIF) in Japan.

Delivery of a healthy newborn using vitrified zygotes that developed from in vitro matured oocytes retrieved from a patient with polycystic ovarian syndrome

The aim of this study is to evaluate vitrified zygotes, which were developed from in vitro matured oocytes that were retrieved from a patient with polycystic ovarian syndrome. Oocyte retrieval was performed on day 15 following withdrawal bleeding. The oocytes were incubated for 24 h in human tissue culture medium (TCM)-199 maturation medium supplemented with 20% filtrated-mixed patients follicle fluid, follicle stimulating hormone (FSH), and human chorionic gonadotropin (hCG). A total of four immature oocytes were collected. Two of the four oocytes (50.0%) developed to the metaphase-II stage and, subsequently, two fertilized oocytes were vitrified at the pronuclear stage because a very thin endometrium was not conducive for transfer. One of the two vitrified zygotes was thawed and developed to a 4-cell cleavage stage embryo within 24 h. Subsequent to the embryo transfer, a healthy newborn was delivered. A successful delivery was ensued by using vitrified zygotes from an anovulatory woman with polycystic ovarian syndrome.

Long-term clinical outcomes of testicular sperm extraction and intracytoplasmic sperm injection for infertile men

To find the best methods to achieve the highest pregnancy and birth rates for couples needing testicular sperm extraction (TESE)‐intracytoplasmic sperm injection (ICSI).

A Review: the Essentials of Informed Consent for Cancer Patients Before Autologous Transplantation of Ovarian Tissue

Abstract: Purpose: To highlight the imperative of informed consent in the fertility preservation of cancer patients before ovarian tissue autotransplantation. Methods: Three papers of tumor recurrence after autotransplantation of frozen-thawed ovarian tissue were compared with the main papers before tumor recurrence was reported in the cancer patients. Results: Histology was performed before autotransplantation in cases 1 and 3, but not in case 2. Histology alone is insufficient for the detection of minimal residual disease (MRD). Furthermore, transplanted ovarian tissue is different from that examined for MRD detection, and how much of the resected ovarian tissue was examined for MRD detection is unclear. The possibility that grafted tissue caused tumor recurrence cannot be ruled out in any of the three cases, because autotransplantation in cancer patients, at present, uses ovarian tissue different from that examined for MRD. Conclusions: We recommend informed consent for cancer patients, because: 1) the transplanted ovarian tissue is different from the ovarian tissue examined for MRD detection; 2) the amount of resected ovarian tissue analyzed for MRD is very small; and 3) MRD detection methods vary. In conclusion, freezing and storage of ovaries should be encouraged, transplantation must be performed carefully, and informed consent is essential.

A successful pregnancy and delivery outcome for a 46-year- old woman following in vitro fertilization

BackgroundDecreased conception rate with advanced maternal age has been well demonstrated. Although this decline becomes apparent at 35 years of age, and accelerates rapidly in women over 40 years of age, we are able to report pregnancy and delivery case for a 46-year-old within vitro fertilization and embryo transfer (IVF-ET). We report a rare case of a successful delivery of a healthy infant by a 46-year-old women following IVF-ET.MethodsShe tried three times with conventional IVF and two times cryopreserved embryo transfer, but she failed to conceive. Low-dose dexamethasone was co-treated the last two times with IVF on the first day of the beginning menstrual cycle daily and continued until retrieval day. We obtained good quality embryos and she became pregnant at the sixth time of IVF which was carried out at the morula and early blastocyst stage.ResultShe finally conceived following co-treatment with low-dose dexamethasone. She delivered vaginally a single male infant with no congenital abnormalities at 40 weeks’ (3416 g; 46, XY).ConclusionWe have demonstrated a successful pregnancy due to obtaining quality embryos using co-treatment low-dose dexamethasone and delivery outcome for a 46-year-old woman using IVF.