Status of online sperm donation and sperm bank in Japan.

Facilities worldwide offering donor sperm insemination face sperm donor shortages. There are also known dangers of online information about sperm donation and using donated sperm to conceive without the involvement of medical professionals. Therefore, the authors investigated 140 Web sites retrieved by common Japanese search engines using sperm donation-related keywords. The authors assessed them according to criteria, but most were deemed unsafe (96.4%). Ultimately, 2 personal and 3 company Web sites provided adequate information. However, the personal Web sites lacked representative individual information and the company Web sites had issues including high costs, overseas location of facilities performing insemination, and possible non-Japanese origin of donated sperm.

Preclinical evaluation of a new cryopreservation container for a limited number of human spermatozoa.

The aim of this study was to develop a new container for cryopreservation of a limited number of spermatozoa. To evaluate the efficacy and safety of this new container, we performed preclinical evaluations using human sperm or mouse oocytes and sperm. First, using human sperm that was frozen and then thawed, we demonstrated that the sperm recovery rate using the new container was 96.7% (58/60), which was significantly higher (P < 0.05) than the recovery rate of 21.2% (11/52) when using the Cryotop®. Sperm motility rates were 19.2% (10/52) using the Cryotop® and 35.0% (21/60) using the new container. Second, murine epididymal spermatozoa were divided into three groups: fresh spermatozoa, spermatozoa frozen using a straw, and spermatozoa frozen using the new container. Sperm motility, sperm membrane and DNA integrity, in vitro development of fertilized eggs, and offspring development after embryo transfer were assessed. The motility of freeze-thawed sperm was lower in spermatozoa that were frozen using the new container than in fresh spermatozoa or those that were frozen using a straw. After intracytoplasmic sperm injection, the survival rate was 96.7% (145/150), the 2-cell development rate was 90.3% (131/145), and the blastocyst development rate was 77.2% (112/145), when using the new container. There were no differences in the sperm membrane, DNA integrity, or in the embryo development rates to the blastocyst stage among the different frozen groups. Six offspring were derived from spermatozoa freeze-thawed in the new container, and they developed normally. Thus, the new container allows easy handling of a small number of sperms and minimizes sperm loss during cryopreservation.

Laparoscopically resected Castleman's disease in the pelvic retroperitoneum: A case report.

Castleman's disease is a rare benign disorder of unknown etiology characterized by proliferation of lymphoid tissues. Castleman's disease arising from pelvic retroperitoneum is clinically rare. The present case report describes a rare case of laparoscopically resected Castleman's disease in the pelvic retroperitoneum associated with benign ovarian cyst. A 47-year-old woman, gravida 5, para 3, was referred to to the Department of Obstetrics and Gynecology of Wakayama Medical University with a suspected pelvic tumor. Magnetic resonance imaging revealed that the solid tumor was localized in the retroperitoneal space at the right side of the pelvis. The patient underwent laparoscopic surgery for the resection of the pelvic retroperitoneal tumor, with complete tumor resection. Postoperative pathological examination established the diagnosis of Castleman's disease. The postoperative course was uneventful, with no evidence of local recurrence or systemic disease 6 months after diagnosis.

The Well-of-the-Well system: an efficient approach to improve embryo development.

Transfer of human embryos at the blastocyst stage may offer considerable benefits including an increased implantation rate and a decreased risk of multiple pregnancies; however, blastocyst culture requires an efficient and reliable in-vitro embryo culture system. In this study, the effect of the Well-of-the-Well (WOW) system consisting of microwells formed on the bottom of the culture dish was tested in three mammalian species, including humans. The WOW system resulted in significant improvement when comparing the drops for culture of in-vitro-matured and parthenogenetically activated porcine oocytes, and in-vivo-derived mouse zygotes. In human embryos, using a sibling oocyte design, embryos cultured in WOW developed to the blastocyst stage in a significantly higher proportion than did embryos cultured traditionally (55% in WOW and 37% in conventional culture; P < 0.05). In a separate study, also in human, a total of 48 patients with a cumulative 214 unsuccessful previous IVF cycles were selected for the trials. In subsequent intracytoplasmic sperm injection cycles, oocytes/embryos were cultured individually in the WOW system or in microdrops. Transferable quality blastocyst development (48.9% of cultured zygotes) was observed in the WOW system. Ninety-four blastocysts transferred to 45 patients resulted in clinical pregnancy rates of 48.9%, including nine twin pregnancies, seven single pregnancies, five miscarriages and one ectopic pregnancy. The results indicate that the WOW system provides a promising alternative for microdrop culture of mammalian embryos, including human embryos.

Stimulation of human damaged sperm motility with hydrogen molecule.

BACKGROUND: Sperm motility is a critical factor in male fertility. Low motility can be caused by a variety factors including abnormal spermatogenesis, oxidative damage, or depletion of intracellular ATP. Recent findings indicate that hydrogen molecule (H2) selectively reduces toxic reactive oxygen species. In this study, we investigated the effects of H2 on human sperm motility in vitro. METHODS: Experimentally damaged sperm suspensions from patients left at room temperature for > 5 days or frozen immediately after ejaculation were used. After exposure with H2, their forward motility was measured with a counting chamber. A time-lapse movie was recorded to analyze sperm swimming speed. Mitochondria were stained with a membrane potential-sensitive dye. RESULTS: H2 treatment significantly improved the rate of forward motility, whereas treatment with nitrogen gas did not. While treatment for 30 min was sufficient to improve motility, it did not affect sperm swimming speed. After 24 h, retreatment with H2 increased the motility again. H2 treatment also increased mitochondrial membrane potential. Forward motility of low motile frozen-thawed sperm from patients significantly improved with cleavage medium containing H2. CONCLUSIONS: Our results illustrated that H2 treatment stimulates low sperm motility. H2 is a new promising tool for male infertility treatments.

Improved postimplantation development of rabbit nuclear transfer embryos by activation with inositol 1,4,5-trisphosphate.

Cloned rabbit embryos are characterized by their extremely poor postimplantation development, despite their high survivability until the blastocyst stage in vitro. This study examined whether the developmental failure of cloned rabbit embryos in vivo can be overcome by technical improvements to the activation protocol. Freshly collected cumulus cells were transferred into enucleated oocytes by intracytoplasmic injection. One to two hours later, the oocytes were activated by electroporation with Ca(2+) or inositol 1,4,5-trisphosphate (IP3), which is known to induce repeated rises in intracellular Ca(2+), as in normal fertilization. After transfer of embryos at the two- to four-cell stages, well-defined implantation sites with remnant fetal tissue were observed at term (day 28) only in the IP3-stimulation groups (0.9% and 5.8% per transferred embryo for single and triple stimulation groups, respectively). When some recipients in the same group were examined at days 16-20, a viable cloned fetus (day 19) with normal organogenesis was obtained. These findings clearly demonstrate that the oocyte activation protocol using IP3 enhances the postimplantation development of nuclear-transferred rabbit embryos.

Production of the first offspring from oocytes derived from fresh and cryopreserved pre-antral follicles of adult mice.

Although mammalian ovaries contain hundreds of thousands of pre-antral follicles, fewer than 1% of these reach maturity and ovulation. Obtaining immature eggs from the pre-antral follicles of ovarian tissue could increase the possibility of preserving fertility in women undergoing anti-cancer treatment, and in women who wish to delay pregnancy and child raising until they are older. This study reports the birth of 10 healthy mouse pups derived from oocytes obtained from pre-antral follicles after adult ovary tissue cryopreservation and allotransplantation. High in-vitro maturation (55.1%), fertilization (76.3%) and cleavage (98.3%) rates were achieved using these oocytes, and there was no significant difference between the vitrified and control samples except in maturation rate (55.1 versus 72.8%, P < 0.05). After an ultra-rapid vitrification procedure, the warmed tissue fragments were transplanted beneath the kidney capsule of severe combined immunodeficient mice for onward in-vivo culture. Within 10 days of culture, 138 full size oocytes developed from the 456 transplanted pre-antral follicles. In-vivo growth of follicles was followed by in-vitro oocyte maturation, in-vitro fertilization and subsequent embryo transfer, leading to the birth of 10 healthy pups. These results may lead to increasing the possibility of preserving fertility by cryopreservation of ovarian tissue.