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Fertilization and development of a blastocyst-stage embryo after selective intracytoplasmic sperm injection of a mature oocyte from a binovular zona pellucida: a case report.

PURPOSE: Our purpose is to describe the development of a blastocyst-stage embryo after the selective fertilization of a mature oocyte from a binovular zona pellucida by intracytoplasmic sperm injection (ICSI). METHOD: A 34-year-old woman underwent intracytoplasmic sperm injection due to severe male-factor infertility. After oocyte retrieval, a binovular zona pellucida including one mature metaphase II oocyte and one immature oocyte at the germinal vesicle stage as well as nine metaphase II oocytes was injected with spermatozoa using a one-to-one approach. RESULTS: The injected mature oocyte of the binovular zona pellucida showed fertilization as evidence by the presence of two pronuclei and cleaved to a four-cell embryo on the second day, while the uninjected oocyte showed signs of degeneration. On the third day, this embryo further cleaved to six blastomeres with slight fragmentation and it reached the blastocyst stage on the sixth day. CONCLUSIONS: Selective fertilization of one oocyte from a binovular zona pellucida by ICSI may lead to the development of a morphologically normal blastocyst-stage embryo which can be used for embryo transfer in the presence of a limited number of embryos.

Adult↗

In vitro fertilization in patients with ovarian endometriomas.

OBJECTIVE: The objective of the study was to establish whether operative treatment of recurrent ovarian endometriosis improves the prognosis of in vitro fertilization. METHODS AND MATERIAL: A retrospective analysis of one hundred endometriosis patients admitted to Tampere University Hospital for IVF treatment. Forty-five patients had an ovarian endometrioma during IVF treatment, 36 of the cases being recurrences after a previous operation. Fifty-five patients had ovarian endometriomas operated without recurrence. The patient groups with or without endometriosis did not differ in age, duration of infertility, sperm parameters, amount of gonadotropins required per oocyte and number of retrieved oocytes. RESULTS: The patients with ovarian endometriosis had more embryos (mean 3.9) than women without endometriomas (mean 2.8) (p<0.05) and the respective pregnancy rates per IVF cycle were 38% and 22%. Patients with endometriomas had a live birth rate of 27% compared with 20% in women with no endometriomas. CONCLUSIONS: The presence of a small endometrioma does not reduce the success of IVF treatment.

Adult↗

Case report: delirium associated with ovarian hyperstimulation syndrome.

Ovarian hyperstimulation syndrome (OHSS) is one of the most important complications of assisted reproductive technologies. Mild OHSS is characterized by ovarian enlargement and abdominal discomfort. In severe cases anasarca, hepatic dysfunction, reduced blood volume, electrolyte imbalance, organ failure and thromboembolic phenomena may be observed. Delirium is a syndrome, not a disease, and has many causes, all of which result in a similar pattern of signs and symptoms relating to a patient's level of consciousness and cognitive impairment. Delirium remains an under-recognized and under-diagnosed clinical disorder. The case is presented of a 30-year-old woman with OHSS and delirium. She underwent intracytoplasmic sperm injection (ICSI) for severe male factor infertility. Five days after oocyte retrieval, ascite formation was observed in ultrasonographic evaluation, and embryo transfer was cancelled. Twelve days after retrieval she came to the emergency clinic with abdominal distension and pain. She was hospitalized and paracentesis was performed every other day three times. She had altered consciousness and psychomotor hypoactivity 1 h after the last paracentesis. Psychiatric consultation revealed that she was in a state of delirium, and haloperidol was administered for treatment. Her symptoms disappeared within a week. Her medication was stopped when symptoms resolved and she was still asymptomatic in psychiatric evaluation 1 week later.

Adult↗

Male smokers have a decreased success rate for in vitro fertilization and intracytoplasmic sperm injection.

OBJECTIVE: Smoking by one or both partners can adversely affect IVF outcome. We investigated whether smoking may also play a role in the success rate of intracytoplasmic sperm injection (ICSI), in which initial steps of fertilization are bypassed. DESIGN: Three hundred one couples (ICSI: 153, IVF: 148) participated in 415 treatment cycles (ICSI: 202, IVF: 213). One hundred thirty-nine men were habitual smokers (ICSI: 71, IVF: 68). Seventy-seven women were smokers (ICSI: 41, IVF: 36). Multiple nominal regression analyses of various steps of assisted reproduction included smoking status, age, semen parameters, and number of embryos transferred. SETTINGS: Reproductive and andrology unit of the university. PATIENT(S): Three hundred one couples seeking fertility treatment. INTERVENTION(S): Assisted reproduction by in vitro fertilization (IVF) or ICSI. MAIN OUTCOME MEASURE(S): Clinical pregnancy. RESULT(S): Intracytoplasmic sperm injection success (clinical pregnancy) in women with smoking male partners was 22% and was 38% with nonsmoking partners. Similar results were seen for IVF, with 18% vs. 32%. Multinominal logistic regression analysis revealed smoking in men to be a significant predictor of ICSI outcome, along with female age and the number of embryos transferred, whereas clinical pregnancies after IVF were dependent on smoking in men, number of embryos transferred, sperm motility, and female age. Female smoking influenced the number of oocytes retrieved and the fertilization rate of oocytes in IVF but not in ICSI. The odds ratio for failure of ICSI for male smokers in comparison to male nonsmokers was 2.95 (IVF: 2.65). CONCLUSION(S): Smoking by males decreases the success rates of assisted reproduction procedures, not only in IVF, but also in ICSI. Apart from putative adverse effects during fertilization, altered DNA in spermatozoa might hamper development of the embryo.

Female↗

Incubation with sperm enhances in vitro maturation of the oocyte from the germinal vesicle to the M2 stage.

OBJECTIVE: To evaluate the effect of sperm in the culture medium on the rate of oocyte maturation in vitro from the germinal vesicle to the M2 stage. DESIGN: Prospective randomized controlled study. SETTING: The IVF Unit, Wolfson Medical Center, Holon, Israel. PATIENT(S): All women in whom oocytes were retrieved at the germinal vesicle stage between December 1995 and March 1996. INTERVENTION(S): Oocytes retrieved at the germinal vesicle stage were divided prospectively and randomly into four groups of incubation conditions: group 1, intact germinal vesicle with cumulus; group 2, intact germinal vesicle with sperm cells in the culture medium; group 3, stripped germinal vesicle; and group 4, stripped germinal vesicle with sperm cells. Oocytes were observed 24 hours after retrieval, and the stage of maturation was recorded. Oocytes that reached the M2 stage underwent the intracytoplasmic injection procedure, and the fertilization rate in each group was recorded at 48 hours. MAIN OUTCOME MEASURE(S): Maturation rate from the germinal vesicle to M2 stage and fertilization rate. RESULT(S): Each group contained 20 germinal vesicle oocytes. In groups 1 and 2, 2 (10%) and 9 (45%) oocytes, respectively, reached the M2 stage at 24 hours; at 48 hours, 1 (5%) and 8 (40%) embryos developed, respectively. The results in group 2 were significantly higher than in group 1. In groups 3 and 4, 6 (30%) and 16 (80%) oocytes, respectively, reached the M2 stage at 24 hours; at 48 hours, 5 (25%) and 14 (70%) embryos developed, respectively. Results in group 4 were significantly higher than those in groups 1, 2, and 3. CONCLUSION(S): Both methods of oocyte activation (i.e., addition of sperm to the culture medium or removal of the cumulus) enhance oocyte maturation in vitro, but the sperm-incubation method has a more pronounced effect. A combination of both methods leads to an exceptionally high rate of oocyte maturation, followed by a high fertilization rate.

Adult↗

[Triplet pregnancy after intracytoplasmic sperm injections].

Morphology of the spermatozoa is one of the major criteria in severe male subfertility. We report on a case of severe Oligoasthenoteratozoospermia in which a triplet pregnancy could be achieved after intracytoplasmic sperm injection (ICSI). After treatment with GnRH-analogues/HMG/HCG seven oocytes of metaphase II could be retrieved and injected. Three of them fertilised and were transferred as four-cell stage embryos after two days. Seven weeks later an intact triplet pregnancy could be detected by ultrasonography. The severe sperm defects in this case were evaluated with the technique of electron microscopy. Ultramorphology of spermatozoa appears to have an important predictive value for fertilisation and pregnancy in classical IVF. Its value for the assisted procreation techniques such as ICSI remains to be investigated.

Adult↗

Pelvic abscess in the second half of pregnancy after oocyte retrieval for in-vitro fertilization: case report.

We describe a very late manifestation of pelvic abscesses after oocyte retrieval for in-vitro fertilization (IVF). In a twin pregnancy achieved after intracytoplasmic sperm injection, rupture of bilateral ovarian abscesses occurred at the end of the second trimester. An emergency laparotomy was necessary because of an acute abdomen. This complication led to severe maternal and neonatal morbidity, preterm birth and neonatal death. The rare occurrence of acute abdomen in pregnancy due to pelvic infection and the non-specific symptoms of a pelvic abscess after oocyte retrieval for IVF are discussed.

Abdomen, Acute↗

Failed fertilization: is it predictable?

PURPOSE OF REVIEW: The purpose of this review is to critically examine the ability of screening tests to predict fertilization failure. RECENT FINDINGS: Failed fertilization occurs in 5-10% of in-vitro fertilization cycles and 2-3% of intracytoplasmic sperm injection cycles. Failed fertilization may result from impaired spermatozoa, oocyte deficiencies or defects in the in-vitro sperm/oocyte medium. In the in-vitro fertilization setting most cases are caused by male factor deficiencies, whereas failure of oocyte activation is the most common cause of failed fertilization after intracytoplasmic sperm injection. Although the standard semen analysis has limited ability to predict fertilization failure, strict sperm morphology criteria, sperm-zona binding ratios and zona pellucida induced acrosome reaction tests provide increased capacity to avoid this outcome. The quality of the semen sample on the day of oocyte retrieval and fertilization performance in previous in-vitro fertilization cycles may also guide the appropriate use of intracytoplasmic sperm injection. However, the routine use of the latter technique in cases of non-male factor infertility is contraindicated. SUMMARY: The ever improving techniques highlighted in this analysis offer improved ability to predict failed fertilization.

Female↗

Successful monozygotic twin delivery following in vitro maturation of oocytes retrieved from a woman with polycystic ovary syndrome: case report.

The incidence of monozygotic twinning (MZT) appears to be increasing within the field of assisted reproductive technology (ART), although the factors contributing to the phenomenon are still far from being identified. On the contrary, in vitro maturation (IVM) of oocytes is becoming more accepted and more and more babies have been born worldwide using this procedure. Assessing its safety and impact on monozygotic twinning (MZT), and following up the health of these babies, is essential. We report here a first case of successful monozygotic (MZ) twin delivery following IVM. The patient was a 28-year-old Japanese female, referred to the IVF clinic for primary infertility. Several previous cycles of ovarian stimulation had resulted in ovarian hyperstimulation syndrome (OHSS). The patient received norethisterone-mestranol to initiate the menstruation, and oocyte retrieval was performed 36 h after hCG. A total of 22 immature oocytes were obtained. Following incubation for 24 h in IVM medium, 50% of the oocytes were matured to the metaphase II (MII) stage. Nine oocytes were fertilized after ICSI with the husband's sperm. Three day 3 embryos were transferred into the uterus on the fourth day following oocyte retrieval. Three weeks after embryo transfer, a single gestational sac was visualized in the uterus. At 7 weeks of gestation, two fetal poles with cardiac activity were seen in the single gestational sac. Serial ultrasound examinations revealed a MZ, monochorionic diamniotic pregnancy. After intensive perinatal monitoring, two healthy male infants were delivered by Caesarean section at 35 weeks of gestation.

Adult↗

Developmental competence of transported in-vitro matured macaque oocytes.

This study examines in-vitro maturation (IVM) in a non-human primate model, Macaca fascicularis. The animals had hormonal injections and laparoscopic oocyte retrieval (OR)) at 12- and 24- h after human chorionic gonadotrophin (HCG). The immature oocytes were placed in tightly capped tubes containing pre-equilibrated IVM medium and transported for 5 h in a dry portable 37 degrees C incubator without CO2 supplement. Meiotic spindle was observed at 36-38- h post-HCG by polarized microscopy in 72 and 84.5% of mature oocytes collected at 12- and 24- h post-HCG oocyte retrieval intervals respectively. However, abnormal spindle formations were detected in some IVM oocytes by confocal microscopy. The IVM oocytes were also randomly selected for (i) intracytoplasmic injection with frozen-thawed epididymal M. fascicularis spermatozoa and (ii) nuclear transfer (NT) with fresh M. fascicularis cumulus cells. Embryonic development of sperm-injected embryos was not affected by the 12- and 24- h post-HCG oocyte retrieval intervals (22.5 versus 27.9% respectively). However, embryonic development of NT embryos was significantly affected by the 12- h post-HCG oocyte retrieval interval (4.5 versus 31.7% respectively; P < 0.01). In conclusion, IVM of monkey oocytes in a dry portable incubator for 5 h did not affect the maturation rate. However, the ability of primate oocytes to develop after somatic cell nuclear transfer was affected by oocyte retrieval time post-HCG.

Animals↗

Outcomes in couples undergoing ICSI: comparison between fresh and frozen-thawed surgically retrieved spermatozoa.

We retrospectively evaluated the impact of cryopreservation on spermatozoa obtained from patients with azoospermia and used for intracytoplasmic sperm injection (ICSI). Frozen-thawed epididymal spermatozoa (FTEPS) was used in 34 couples, whereas frozen-thawed testicular spermatozoa (FTTS) was used in 50 couples for ICSI during assisted conception, and these results were compared with results using fresh spermatozoa for ICSI in the same individuals. The fertilization rate (FR) was significantly lower for FTTS (65.8%) but not for FTEPS (73.1%) compared with the FR using fresh spermatozoa (72.3% and 73.2% respectively). In contrast, neither the implantation nor the pregnancy rate was altered when FTEPS or FTTS was used. In conclusion, our results indicate that surgically retrieved spermatozoa can be efficiently used for ICSI after freezing and thawing without compromising the outcome.

Cryopreservation↗

Fertilization and pregnancy outcome with intracytoplasmic sperm injection for azoospermic men.

The evident ability of the intracytoplasmic sperm injection (ICSI) procedure to achieve high fertilization and pregnancy rates regardless of semen characteristics has induced its application with spermatozoa surgically retrieved from azoospermic men. Here, ICSI outcome was analysed in 308 cases according to the cause of azoospermia; four additional cycles were with cases of necrozoospermia. All couples were genetically counselled and appropriately screened. Spermatozoa were retrieved by microsurgical epididymal aspiration or from testicular biopsies. Epididymal obstructions were considered congenital (n = 138) or acquired (n = 103), based on the aetiology. Testicular sperm cases were assessed according to the presence (n = 14) or absence (n = 53) of reproductive tract obstruction. The fertilization rate using fresh or cryopreserved epididymal spermatozoa was 72.4% of 911 eggs for acquired obstructions, and 73.1% of 1524 eggs for congenital cases; with clinical pregnancy rates of 48.5% (50/103) and 61.6% (85/138) respectively. Spermatozoa from testicular biopsies fertilized 57.0% of 533 eggs in non-obstructive cases compared to 80.5% of 118 eggs (P = 0.0001) in obstructive azoospermia. The clinical pregnancy rate was 49.1% (26/53) for non-obstructive cases and 57.1% (8/14) for testicular spermatozoa obtained in obstructive azoospermia, including three established with frozen-thawed testicular spermatozoa. In cases of obstructive azoospermia, fertilization and pregnancy rates with epididymal spermatozoa were higher than those achieved using spermatozoa obtained from the testes of men with non-obstructive azoospermia.

Biopsy↗

Analysis of meiosis in intratesticular germ cells from subjects affected by classic Klinefelter's syndrome.

Azoospermic subjects affected by Klinefelter's syndrome may occasionally show the presence of intratesticular residual foci of spermatogenesis, and the retrieval of mature spermatozoa from the testis may permit fertility and paternity by means of intracytoplasmic sperm injection. Previous studies have demonstrated that these subjects show the presence of an increased incidence of hyperaploid spermatozoa. Here we analyzed, by fluorescence in situ hybridization using specific probes for chromosomes 8, X, and Y, the spermatogenic process and the meiotic progression of 47,XXY germ cells retrieved by fine needle aspiration of the testis in ten azoospermic patients affected by classic Klinefelter's syndrome. All patients had lower testicular volume, higher gonadotropins, and lower testosterone plasma levels compared with control subjects. Cytological analysis of the testicular cells retrieved by fine needle aspiration showed the presence of Sertoli cells only in eight subjects, while germ cells were observed in two patients. In each patient Sertoli cells showed a 47,XXY karyotype, and the same chromosome pattern was observed in spermatogonia and primary spermatocytes of patients presenting a residual spermatogenesis. Secondary spermatocytes, spermatids, and mature spermatozoa showed different sex chromosome patterns, reflecting their origin from 47,XXY spermatogonia. In conclusion, this study demonstrated that, in subjects affected by Klinefelter's syndrome, residual germ cells may be present in the testis and that 47,XXY spermatogonia are able to undergo and complete the spermatogenic process leading to mature spermatozoa. These data further suggest the need to evaluate the sex chromosome status of sperm from patients affected by Klinefelter's syndrome undergoing assisted reproductive techniques.

Adult↗

A survey of four years of experience with intracytoplasmic sperm injection.

The present report covers the results of a 38-month period in which 2853 consecutive intracytoplasmic sperm injection (ICSI) cycles were performed in 1953 couples. These couples were afflicted with male factor infertility and had at least one previous failed conventional in vitro fertilization (IVF) treatment cycle. In other couples, the husband had semen parameters incompatible with conventional IVF or suffered from excretory or secretory azoospermia where it was possible to recover spermatozoa by microsurgical epididymal sperm aspiration (mesa) or by testicular sperm extraction (tese) procedure. Overall, the 2-PN fertilization rate was 62% per retrieved metaphase II oocyte and 70% per successfully injected metaphase II oocyte. Embryo transfer was performed in 91% of started cycles. The cumulative pregnancy rate (positive HCG) was 34% per started ICSI treatment and 37% per embryo transfer.

Adult↗

Dizygotic twin delivery following in vitro fertilization and transfer of thawed blastocysts cryopreserved at day 6 and 7.

OBJECTIVE: To report the first conception and delivery following transfer of thawed human blastocysts maintained in extended in vitro culture with cryopreservation at day 6 and 7. DESIGN: Case report. SETTING: Major urban infertility referral center. PATIENT(S): A 26-year-old woman with pelvic endometriosis and two prior unsuccessful in vitro fertilization/embryo transfer (IVF-ET) attempts. INTERVENTION(S): The patient underwent controlled ovarian hyperstimulation using a combined FSH + hMG protocol, and 24 oocytes were retrieved. MAIN OUTCOME MEASURE(S): Dizygotic twin delivery after IVF and intracytoplasmic sperm injection (ICSI), assisted embryo hatching, and ultrasound-guided transfer of cryopreserved blastocysts. RESULT(S): After three embryos were subjected to assisted hatching, they were transferred fresh on day 3, but no implantation occurred. All nontransferred embryos (n = 11) were observed during extended in vitro culture and three blastocysts were selected for cryopreservation on day 6 and 7; thaw and transfer occurred the following month and a pregnancy was achieved. Dizygotic twins (female/female) were delivered by cesarean in the early third trimester. CONCLUSION(S): Substantial advancements have been made in the field of embryo cryogenics and in vitro fertilization, but controversy remains regarding the value of freezing late-developing human blastocysts. Here we describe the first reported live births with IVF after extended in vitro culture and cryopreservation at day 6 and 7 after fertilization.

Adult↗

Orthotopic autotransplantation of cryopreserved ovarian tissue to a woman cured of cancer - follicular growth, steroid production and oocyte retrieval.

Cryopreservation of human ovarian tissue is now an option for cancer patients facing treatment with gonadotoxic regimes, as a means of preserving their fertility. So far, there have been only a few reports on autotransplantation of frozen-thawed tissue with regard to restoration of ovarian function. The present report describes a 32-year-old woman diagnosed with Hodgkin's lymphoma, who had cryopreserved ovarian tissue transplanted orthotopically after secondary ovarian failure due to chemotherapy. Only 8 weeks after transplantation, ultrasonography of the remaining ovary revealed two follicles with diameters of 10 and 15 mm. Concomitantly, circulating concentrations of oestradiol increased, while concentrations of gonadotrophins decreased. In the following months, the patient menstruated three times. Subsequent pituitary down-regulation with a gonadotrophin-releasing hormone (GnRH) agonist and ovarian stimulation resulted in development of one pre-ovulatory follicle from which a metaphase II oocyte was retrieved; however, this oocyte was unable to sustain further development after intracytoplasmic sperm injection (ICSI). Intrafollicular concentrations of oestradiol and progesterone suggested a normal luteinizing response of the follicle to human chorionic gonadotrophin stimulation. A 7-month follow-up revealed continued vivid follicular activity and normal oestradiol concentrations. In conclusion, cryopreserved human ovarian tissue restored ovarian function for several cycles and sustained development of mature oocytes in a woman cured of cancer.

Adult↗

High doses of gonadotrophin-releasing hormone antagonist in in-vitro fertilization cycles do not adversely affect the outcome of subsequent freeze-thaw cycles.

The clinical application of gonadotrophin-releasing hormone (GnRH) antagonists instead of GnRH agonists, to prevent spontaneous premature luteinizing hormone surge during ovarian stimulation for assisted reproduction treatment has been advocated. A recent, double-blind, dose-finding study, including six dosages of the GnRH antagonist ganirelix, in women undergoing ovarian stimulation with recombinant follicle stimulating hormone (FSH), has indicated that high doses of GnRH antagonist (1 or 2 mg once daily) are associated with a low implantation rate. This follow-up study reports on the pregnancy rate after replacement of cryopreserved embryos obtained in stimulation cycles of the above-mentioned trial. Ovarian stimulation was initiated on day 2 of the cycle, with daily injections of 150 IU recombinant FSH. Ganirelix (0.0625, 0.125, 0.25, 0.5, 1.0 or 2.0 mg) was administered once daily from stimulation day 6 onwards, up to and including the day of human chorionic gonadotrophin. Retrieved oocytes were fertilized by in-vitro fertilization (IVF) or intracytoplasmic sperm injection and a maximum of three fresh embryos was transferred. Excess embryos were frozen, and subsequently used in either natural or programmed cycles. Until June 1998, 11 ongoing pregnancies (12-16 weeks after embryo transfer) were achieved from 46 cycles in which embryos had been first frozen (23.9% per transfer). Six of these 11 patients had been treated with a high dose of ganirelix (1.0 or 2.0 mg) during the IVF cycles in which the embryos were obtained. In conclusion, our data suggest that high dosages of ganirelix do not adversely affect the potential of embryos to establish clinical pregnancy in freeze-thaw cycles.

Adult↗

Microsurgical TESE and the distribution of spermatogenesis in non-obstructive azoospermia.

We wished to map the distribution of spermatogenesis in different regions of the testis in 58 men with non-obstructive azoospermia, and to develop a rational microsurgical strategy for the testicular sperm extraction (TESE) procedure. One goal was to maximize the chances for retrieving spermatozoa from such men, to minimize tissue loss and pain, and to preserve the chance for successful future procedures. Another goal was to expand upon the previously reported quantitative histological analysis of testicular tissue in 45 azoospermic men undergoing conventional TESE, this time using microsurgical as well as histological mapping. Tubular fullness observed at microsurgery and the presence of spermatozoa in the TESE specimen was compared with the quantitative histological analysis of spermatogenesis. Thus, our conclusions about the distribution of spermatogenesis are based on our experience with TESE in 103 consecutive cases of non-obstructive azoospermia. It was confirmed that men with non-obstructive azoospermia caused by germinal failure have a mean of 0 to 3 mature spermatids per seminiferous tubule in contrast to 17-35 mature spermatids per tubule in men with normal spermatogenesis and obstructive azoospermia. The former represented the threshold of quantitative spermatogenesis which must be exceeded in order for spermatozoa to 'spill over' into the ejaculate. Both testicular 'mapping' by multiple biopsy (n = 15) and microsurgical removal of contiguous strips of testicular tissue (n = 43) revealed a diffuse, rather than regional, quantitative distribution of spermatogenesis. A microsurgical approach resulted in the minimal amount of tissue loss and minimal-to-no pain (compared with the original 45 cases already reported). By this means it is often possible to immediately locate the few tubules with spermatogenesis at microsurgery, under local anaesthesia. But even in cases where greater amounts of tissue must be removed in order to find spermatozoa, the microsurgical TESE procedure prevents secondary testicular damage by protecting blood supply and preventing pain and atrophy from increased testicular pressure. Thus, future attempts at TESE-ICSI need not be compromised.

Biopsy↗