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

Purpose: To find the best methods to achieve the highest pregnancy and birth rates for couples needing testicular sperm extraction (TESE)-intracytoplasmic sperm injection (ICSI). Methods: Retrospectively 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). Results: The 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. Conclusion: Intracytoplasmic 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.

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

Aim: 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. Methods: Fifty patients with a history of RIF with frozen-thawed blastocyst transfers were recruited from July, 2015 to April, 2016. Endometrial sampling for the ERA and histological dating and a pET according to the ERA were performed. The receptive (R) or non-receptive (NR) status of the endometrium as a result of the first ERA, endometrial dating, and pregnancy rates after the pET were analyzed. Results: Of the patients with RIF, 12 (24%) were NR. Among them, eight (66.7%) were prereceptive. A clinical follow-up was possible in 44 patients who underwent the pET. The pregnancy rates were 58.8% per patient and 35.3% per first pET in the R patients and 50.0% per patient and 50.0% per first pET in the NR patients. Discrepancies between the ERA results and histological dating were seen more in the NR patients than in the R patients. Conclusions: For patients with unexplained RIF, there is a significance in searching for their personal window of implantation (WOI) using the ERA, considering the percentage of those who were NR and the pregnancy rates that resulted from the pET. By transferring euploid embryos in a personal WOI, much better pregnancy rates are expected.

Identification of new risk factors for wound separation in gynecologic malignancy surgery.

AIM: The purpose of this study was to compare prophylactic subcutaneous drainage plus subcuticular sutures versus staples for the risk of wound separation after skin closure following gynecologic malignancy surgery, and to investigate the risk factors of this procedure. MATERIAL AND METHODS: Patients were divided into two groups: 120 patients who were treated with subcutaneous drainage plus subcuticular sutures (Suture group) and 201 patients with staples plus subcutaneous sutures (Staples group). In the Suture group, subcuticular tissue was approximated with interrupted 4-0 polydioxanone sutures, and adhesive closure strips were used on the skin surface. A 3.3-mm closed drainage was implicated in subcutaneous tissue. In the Staples group, subcutaneous tissue was approximated with interrupted polyglactin (Vicryl, Ethicon) sutures. RESULTS: Baseline characteristics were not significantly different between the two groups. Mean operation times were compatible (201 vs 196 min, P = 0.16). The incidence of wound separation was less in the Suture group than in the Staples group (3/120 vs 17/201, P = 0.033). Multiple logistic regression analysis revealed that the Staples group was an independent risk factor for wound separation (odds ratio 7.34, 95% confidence interval: 1.59-33.91, P = 0.011), independent of obesity, International Federation of Gynecology and Obstetrics stages, and operation time. None of the 14 obese patients in the Suture group showed surgical wound separation. CONCLUSIONS: The combination of a prophylactic subcutaneous drain and subcuticular sutures reduced wound separation after skin closure following gynecologic malignancy surgery. With the information regarding risk factors established in this study, the above method provides the best results to minimize the risk, particularly in obese patients.

Deliveries of babies with normal health derived from oocytes with smooth endoplasmic reticulum clusters.

PURPOSE: To examine the impact on development of derived embryos from smooth endoplasmic reticulum clusters (SERC) in human metaphase II (MII) oocytes. METHODS: Retrospective analysis at Kyono ART Clinic. Comparison of embryological development, pregnancy, live birth and fetal malformation between oocytes with SERC (the SERC(+) group) and those without (the SERC(-) group) in 2,158 patients (3,758 cycles) after ICSI. RESULTS: Fertilization and implantation rate were significantly lower in SERC(+) MII oocytes than in SERC(-) MII oocytes. After the transfer of fresh and vitrified embryos derived from SERC(+) oocytes, 14 pregnancies resulted in 14 healthy babies, including 2 from fresh embryo transfer (ET) and 12 from vitrified-warmed ET, with no malformations. CONCLUSION(S): The presence of SERC in MII oocytes was associated with significantly lower fertilization rates and implantation rates than seen in SERC(-) MII oocytes within SERC (+) cycles. However, SERC had no impact on post-implantation development as well as neonatal outcome.

A live birth from vitrified-warmed oocytes in a Philadelphia chromosome-positive acute lymphoid leukemia patient 5 years following allogenic bone marrow transplantation and after a magnitude 9.0 earthquake in Japan.

PURPOSE: To report a live birth from vitrified-warmed oocytes for a Philadelphia chromosome-positive acute lymphoid leukemia (Ph-ALL) patient. METHODS: A 20-year-old single woman with Ph-ALL requested oocyte cryopreservation at a private fertility clinic using assisted reproduction technology (ART). In cases of leukemia, there is a very short time before chemotherapy, follwed shortly by total body irradiation (TBI), and although she had already received the chemotherapy, ten oocytes were vitrified and stored for 59 months before warming. Soon after the oocyte cryopreservation, she received TBI and bone marrow transplant (BMT). During the storage, a magnitude 9.0 earthquake occurred making oocyte transport necessary. The embryo transfer was planned in a hormone replacement cycle, and intracytoplasmic sperm injection (ICSI) was performed on the vitrified-warmed oocytes. On day 3, two embryos were transferred. RESULTS: The patient became pregnant and delivered a healthy girl after ICSI using vitrified-warmed oocytes. CONCLUSIONS: Oocyte cryopreservation is the best option for fertility preservation of young single women with leukemia. Oncologists and gynecologists who conduct ART should cooperate to improve the quality of life of cancer patients.

Birth of a healthy infant after intracytoplasmic sperm injection using pentoxifylline-activated sperm from a patient with Kartagener's syndrome.

OBJECTIVE: To describe the delivery of a healthy female infant after intracytoplasmic sperm injection (ICSI) using pentoxifylline-activated sperm from a patient with Kartagener's syndrome. DESIGN: Case report. SETTING: Private assisted reproductive technology clinic in Japan. PATIENT(S): A couple with male factor infertility due to Kartagener's syndrome. INTERVENTION(S): Intracytoplasmic sperm injection using ejaculated sperm activated by pentoxifylline. MAIN OUTCOME MEASURE(S): Semen characteristics, sperm ultrastructure, fertilization, pregnancy, and birth after ICSI. RESULT(S): The fertilization rate was 7 of 12 (58.3%), and the blastocyst formation rate was 4 of 7 (57.1%); all blastocysts were vitrified. After a single blastcyst transfer, a pregnancy ensued and progressed to term; a healthy female infant was delivered. CONCLUSION(S): With ejaculated sperm, which was activated by pentoxifylline, successful fertilization was accomplished by ICSI; thus, fertilization, vitrification, pregnancy, and delivery are attainable with sperm obtained from men with Kartagener's syndrome.

A birth of twins-one boy and one girl-from a single embryo transfer and a possible natural pregnancy.

PURPOSE: To describe a rare case of a birth of dizygotic twins with different-sex infants from a single embryo transfer. METHODS AND RESULTS: A patient, who had her right ovary and tube removed, and her husband were treated with ICSI and a single embryo transfer. When a single fresh embryo was transferred on day 4, following oocyte retrieval using GnRH agonist-long protocol, two gestational sacs were recognized at 8 weeks of gestation. Healthy twins with a boy and a girl were delivered at 37 weeks 0 days of gestation by a cesarean section. The boy's weight was 2096g, and his height was 45.0 cm, while the girl's weight was 1988g, and her height was 41.5 cm. Peripheral lymphocyte chromosome analysis of the two infants showed normal karyotype, 46, XY (boy) and 46, XX (girl). CONCLUSIONS: A single embryo transfer could produce different-sex twins.

Birth of a healthy male infant after transfer of vitrified-warmed blastocysts derived from intracytoplasmic sperm injection with vitrified-warmed oocytes and frozen-thawed spermatozoa.

PURPOSE: To report a successful delivery of a healthy baby after transfer of vitrified-warmed blastocysts derived from introcytoplasmic sperm injection (ICSI) with vitrified-warmed oocytes and frozen-thawed sperm. METHODS: A female patient and her husband with non-obstructive azoospermia received a transfer of vitrified-warmed blastocysts from vitrified-warmed oocytes and frozen-thawed sperm. The main outcome measures were fertilization, pregnancy and birth. RESULTS: Nine oocytes were matured and vitrified. When the vitrified oocytes were warmed, six survived with good quality morphology. Using ICSI, frozen-thawed sperm was injected into the six warmed oocytes that survived, and the fertilization rate was 100%. The zygotes were cultured, and five of six early embryos became blastocysts. One of them was transferred, but pregnancy was not achieved. The second time around, two vitrified-warmed blastocysts were transferred resulting in pregnancy, and a healthy boy was delivered. CONCLUSIONS: This is a rare case of a successful birth using a vitrified-warmed blastocyst grown after ICSI with a vitrified-warmed oocyte and frozen-thawed sperm.

A birth from the transfer of a single vitrified-warmed blastocyst using intracytoplasmic sperm injection with calcium ionophore oocyte activation in a globozoospermic patient.

OBJECTIVE: To present the effectiveness of diagnostic heterologous intracytoplasmic sperm injection (ICSI), mouse oocyte activation test (MOAT), and ICSI combined with assisted oocyte activation (AOA) in a globozoospermic patient. DESIGN: A case report. SETTING: A private IVF center, Japan. PATIENT(S): A patient with globozoospermia. INTERVENTION(S): MOAT in a mouse and ICSI combined with AOA in a human. MAIN OUTCOME MEASURE(S): Ultrastructure, MOAT, fertilization, and pregnancy. RESULT(S): The transmission electron micrographs showed 100% round-headed spermatozoa lacking an acrosome. MOAT showed that the fertilization rate was 68.4% (13/19) when AOA was used but 0% (0/19) when AOA was not used. After the diagnosis of globozoospermia and sperm-related activation deficiency, 17 human mature oocytes were activated with calcium ionophore after ICSI was performed. The fertilization rate was 88.2% (15/17), and 11 blastocysts were cryopreserved using the vitrification method to prevent severe ovarian hyperstimulation syndrome. A single vitrified-warmed blastocyst was transferred. A gestational sac with fetal heart movements was recognized, and a healthy boy weighing 3180 g was born at 40 weeks of gestation by cesarean section without any congenital abnormality. CONCLUSION(S): MOAT allows discrimination between sperm- and oocyte-related fertilization failures and shows the effectiveness of AOA.

Birth and follow-up of babies born following ICSI using SrCl2 oocyte activation.

This report describes six successful pregnancies (five healthy children from four deliveries and two miscarriages) with SrC1(2) oocyte activation using spermatozoa from nine patients with repeated fertilization failure. Oocytes were artificially activated by SrC1(2) 30 min after intracytoplasmic sperm injection (ICSI). Oocytes were placed in 10 mmol/l of SrC1(2) medium for 1 h, rinsed several times, and then cultured in Universal IVF medium. Developmental characteristics of five resulting children until 1 year old were assessed according to the maternal and children health hand book issued by Mothers' and Children's Health Organization in Japan. Mean fertilization rate, mean frequency of good cleaved embryos, pregnancy rate, and implantation rate after artificial activation in nine couples were increased from 21.7 to 64.5% (P < 0.001), from 0 to 15.4%, from 0 to 40.0% and from 0 to 25.0% respectively. Five healthy children were born following ICSI and artificial activation between February 2005 and March 2006. Physical and mental development of the children from birth to 12 months was normal. These suggest the utility and safety of SrCl(2) for patients with repeated failed fertilizations following ICSI and artificial activation.

Seven pregnancies and deliveries from non-mosaic Klinefelter syndrome patients using fresh and frozen testicular sperm.

PURPOSE: The aim of this study was to investigate the feasibility of using frozen-thawed testicular sperm as well as the timing of testicular sperm extraction (TESE) in patients with non-mosaic Klinefelter syndrome. METHODS: Intracytoplasmic sperm injection (ICSI) was performed in six of 17 (35%) patients whose sperm was recovered by TESE. Multiple biopsies of both testes were performed on the day of oocyte retrieval in all but one of the six patients. RESULTS: Seven pregnancies and deliveries were achieved in five couples, and one couple was unsuccessful. Five pregnancies were achieved using fresh motile sperm, and two were achieved using frozen-thawed sperm. Sperm cryopreservation was not possible in one of the five couples because of the small number of recovered sperm, and possible in four other couples for subsequent ICSI. One woman whose husband had TESE performed prior to ovarian stimulation did not become pregnant. This may be due to the attainment of only a few immotile sperm following the frozen-thawed procedure. CONCLUSION: The outcome of ICSI using fresh or frozen-thawed testicular sperm in patients with non-mosaic Klinefelter syndrome was identical; however, TESE should be performed on the day of oocyte retrieval until such time as a procedure with a higher sperm yield from TESE is available. Moreover, an improved recovery procedure after cryopreservation-thawing of a single spermatozoon must be developed.

Successful pregnancy and delivery after transfer of a single blastocyst derived from a vitrified mature human oocyte.

OBJECTIVE: To describe the first case of delivery after the transfer of a single blastocyst derived from a vitrified mature human oocyte. DESIGN: Case report. SETTING: Private assisted reproduction clinic. PATIENT(S): A normal 31-year-old woman. INTERVENTION(S): An unsuccessful attempt was made to extract sperm from the patient's nonobstructive azoospermic husband by testicular sperm extraction. MAIN OUTCOME MEASURE(S): Transfer of single blastocyst derived from vitrified human oocyte and donor sperm. RESULT(S): A healthy male neonate weighing 3000 g was born. CONCLUSION(S): Vitrification is a useful method of preserving mature human oocytes and has the advantage of time-effectiveness, simplicity, cost reduction, and no need for devices such as programmable freezers.

Successful pregnancy and delivery following combined treatment of in vitro maturation (IVM) and testicular sperm extraction (TESE): a case report.

PURPOSE: To report the efficacy of the combined treatment of in vitro maturation (IVM) and testicular sperm extraction (TESE). METHODS: A couple in which the wife had polycystic ovarian syndrome and the husband had severe oligozoospermia. Oocytes were cultured in vitro for maturation followed by oocytes pickup with natural cycle, and TESE was undergone for husband. Matured oocytes were fertilized by intracytoplasmic sperm injection, and two embryos were transferred to wife's uterine. RESULTS: This case was achieved during pregnancy and delivery of a healthy female infant. CONCLUSIONS: The combined treatment of IVM and TESE was effective for this couple's specific infertility factors.

Freezing of human immature oocytes using cryoloops with Taxol in the vitrification solution.

PURPOSE: In human frozen immature oocytes, there has been little successful delivery. We examined the feasibility of vitrification solution including Taxol, cytoskeltal stabilizer. METHODS: We set four experimental groups that immature oocytes has cumulus cells or not, or including Taxol or not in the vitrification solution. Frozen-thawed oocytes have been performed IVM, ICSI, and IVC. RESULTS: There were no significant differences in survival, maturation, and fertilization rate, respectively. However, in the group enveloped by cumulus cells and including Taxol in the vitrification solution, one embryo was developed to blastocyst. CONCLUSIONS: Our results showed that using vitrification solution with Taxol proved so effective.

Physical and mental development of children after in vitro fertilization and embryo transfer.

Objective: To evaluate the physical and mental development of children after in vitro fertilization (IVF) and frozen embryo transfer (FET). Methods: Between July 1995 and November 2003, 506 patients delivered 658 babies after IVF and ET treatment at our clinic (intracytoplasmic sperm injection (ICSI), 418; conventional IVF, (C-IVF) 125; FET, 115). A survey of the physical and mental developmental of the children was conducted by mailing questionnaires to parents. Comparisons were made between three treatment procedures, and development of singleton, twin and triplet delivery. Results: The response rate was 73.4% (483/658) for 324 children born after ICSI, 78 born after C-IVF, and 81 born after FET. The height and weight of assisted reproductive technology (ART) children at birth were significantly lower than that of naturally conceived babies (ART children: natural delivery; 46.8 cm, 49.0 cm and 2524 g, 3040 g, respectively). However, there was no significant difference between the singletons alone and naturally conceived children irrespective of the ART method. In addition, mental development was the same between singletons and naturally conceived children. The ART group tended to delay body development such as 'holding their head up', 'sitting up', 'crawl' to moving growth in multiple births. Conclusion: The physical and mental development of twins or triplets was significantly more delayed than that of naturally conceived babies, but had improved to a similar extent of the singletons after the age of 6 months. (Reprod Med Biol 2004; 3: 63-67).

Two-step consecutive transfer of early embryos and blastocysts.

Aim: To evaluate and confirm the merit of two consecutive transfer attempts of early embryos and blastocysts. Methods: A total of 685 patients underwent routine in vitro fertilization (IVF) cycles. The study population consisted of three groups: (i) Group 1, early stage embryos were transferred (460 patients; 567 cycles); (ii) group 2, blastocysts were transferred (88 patients; 105 cycles); and (iii) group 3, a two-step (consecutive) transfer was performed on 137 women (141 cycles). Specifically, a standard embryo transfer was performed on day 3, together with a two-step (consecutive) transfer of blastocysts. After the early embryo transfer, an extended culture of supernumerary embryos was conducted, followed by a second transfer of blastocyst(s). Results: No significant differences were found in the three groups with regards to either pregnancy or implantation rates for groups 1, 2, or 3; pregnancy: 34.6, 29.9, and 33.6%, respectively; implantation: 18.6, 15.9, and 15.1%, respectively. The miscarriage rate for each group was also not significant; 20.4, 30.8 and 28.6% for groups 1, 2 and 3, respectively. The multiple pregnancy rate of the three groups was 30.4, 30.8 and 35.7%, respectively, and these were not found to be significant results. Conclusion: No significant difference was found between the three groups with regards to the pregnancy, implantation, multiple pregnancy and miscarriage rates. The multiple pregnancy rate was highest in the two-step (consecutive) transfer group. This difference was not regarded to be highly significant. (Reprod Med Biol 2003; 2: 133-137).

Successful delivery following cryopreservation of zygotes produced by in vitro matured oocytes retrieved from a woman with polycystic ovarian syndrome-like disease: a case report.

PURPOSE: To estimate frozen zygotes, which developed from in vitro matured oocytes retrieved from polycystic ovarian syndrome-like disease. METHODS: Oocyte retrieval was performed on Day 15 following withdrawal bleeding. The oocytes were incubated for 24 h in TCM-199 maturation medium supplemented with follicle fluid, E2, FSH, and hCG. RESULTS: A total of 12 immature oocytes were collected. Seven of the 12 oocytes (58.3%) developed to the metaphase-II stage, and subsequently, all seven fertilized oocytes were frozen at the pronuclear stage. The remaining five oocytes failed to develop to the metaphase-II stage after an additional 24 h of incubation. Three of seven cryopreserved oocytes were thawed and developed to 2-8-cell cleaved stage embryos. The first pregnancy failed. However, the second frozen-thawed embryo transfer resulted in the delivery of healthy twins. CONCLUSIONS: Successful delivery using frozen zygotes from an anovulatory woman with polycystic ovarian syndrome-like disease.