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.

PurposeTo assess the efficacy of fertility preservation (FP) and the impact of chemotherapy on the reproductive potential of Japanese patients with breast cancer.MethodsSixty‐two patients with breast cancer visited the authors’ centers from October, 2003 to June, 2015. They were divided into two groups according to the treatment: oocyte or embryo vitrification for FP before cancer treatment (group A) or infertility treatment after cancer treatment (group B). Group B was divided into two subgroups, B1 (no chemotherapy) and B2 (postchemotherapy), in order to analyze the effect of anticancer drugs on ovarian reserves and assisted reproductive technology outcomes. The number of retrieved oocytes, vitrified oocytes or embryos, and pregnancy rates were analyzed and compared: group A compared to group B1 compared to group B2.ResultsThe patients in groups A and B1 underwent egg collection without any chemotherapy. The numbers of collected oocytes and vitrified embryos were significantly higher in groups A and B1 than in group B2. Nearly 50% of the in vitro fertilization patients who underwent an embryo transfer (ET) became pregnant, including two patients in group A who underwent a vitrified‐warmed ET. Among the pregnant women, 70% did not have chemotherapy.ConclusionFor patients with breast cancer, FP with unfertilized oocytes or embryos before chemotherapy seems to be promising for achieving higher pregnancy rates, with no risk of minimal residual disease.

This model has been termed the Danish model ("the woman stays - the tissue moves"). This is truly patient-centered medicine. We can have maximum effects with the minimum burden. A transportation network like those of Denmark and Germany is the best strategy for FP in Japan. It may be the best system for cancer patients, medical staff, and the Ministry of Health, Labor, and Welfare.

An experiment was conducted to investigate pH, osmolality, and the concentration of ammonia, total protein, glutamine and glutamic acid in follicular fluid at different developmental stages (<2, 3-4 and 5-6 mm in diameter) and serum of porcine. The concentrations of ammonia and total protein content were determined with the catalyzed indophenols reaction and the Bradford assay method and read on a spectrophotometer set at 625 and 595 nm, respectively. Glutamine and glutamic acid concentrations were determined by HPLC. The pH value was lower (P<0.05) but osmolality was higher (P <0.05) in follicular fluid than in s erum. The concentration of ammonia was lower (P<0.05) in follicular fluid than in serum. On the other hand, glutamine and glutamic acid concentrations were higher (P<0.05) in the follicular fluid than in serum. The pH increased and osmolality decreased with increasing follicle size, and protein content was almost similar in small, medium and large follicular fluid. Ammonia, glutamine and glutamic acid concentrations decreased (P<0.001) as follicular size increased. During early follicular development ammonia and amino acids were synthesized for high metabolic breakdown of protein and gradually decreased due to metabolism of ammonia and glutamic acid to glutamate.

PurposeTo find the best methods to achieve the highest pregnancy and birth rates for couples needing testicular sperm extraction (TESE)‐intracytoplasmic sperm injection (ICSI).MethodsRetrospectively studied were 801 patients with male factor infertility who had undergone TESE‐ICSI between April, 1996 and July, 2016 and who had been categorized into four groups: obstructive azoospermia (OA); non‐obstructive azoospermia (NOA); Klinefelter syndrome (KS); and cryptozoospermia (Crypt). The sperm retrieval rate, hormone levels, fertilization rate (FR), pregnancy rate (PR), and birth rate (BR) after ICSI among three groups were compared: fresh testicular sperm (FS)‐fresh oocytes (FO) (Group I); frozen‐thawed testicular sperm‐FO (Group II); and FS‐vitrified‐warmed oocytes (Group III).ResultsThe testicular sperm recovery rate was 57.8% (463/801): 89.6% in the Crypt, 97.1% in the OA, 28.9% in the NOA, and 42.2% in the KS groups. The follicle‐stimulating hormone levels were significantly higher in the NOA and KS groups and the testosterone levels were significantly lower in the KS group. The FR, PR, and BR were: 65.2%, 43.2%, and 28.5% in group I; 59.2%, 33.4%, and 18.7% in group II; and 56.4%, 33.8%, and 22.1% in group III.ConclusionIntracytoplasmic sperm injection with FS‐FO achieved the best PR and BR. It should be considered what to do in cases with no testicular sperm by TESE. The authors hope that ICSI with donor sperm will be allowed in Japan in the near future.