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Impact of oocyte pre-incubation time on fertilization, embryo quality and pregnancy rate after intracytoplasmic sperm injection.

Although, it is well known that pre-incubation of oocytes prior to conventional IVF improves fertilization and pregnancy rates, there are conflicting results regarding the effect of pre-incubation time in ICSI. This study evaluated the role of pre-incubation of oocytes on outcome in intracytoplasmic sperm injection (ICSI) cycles. A total of 1260 patients undergoing their first ICSI cycles were evaluated retrospectively. In patients undergoing ICSI during the year 2000 (Group I, n = 670), oocytes were injected immediately after retrieval, whereas in patients undergoing ICSI during 2001 (Group II, n = 590), oocytes were incubated for 2-4 h prior to injection. The mean age of patients was 33.9 +/- 5.04 years and 34.1 +/- 5.06 years in groups I and II, respectively. The number of oocytes with a first polar body (MII) and fertilization and cleavage rates were higher, and embryo quality was significantly better in group II. In contrast, the total numbers of oocytes without a first polar body (MI), those where germinal vesicle breakdown had not occurred (GV), and empty zona oocytes were higher in group I. No difference was found in the number of embryos transferred or implantation or clinical pregnancy rates. This study demonstrated that pre-incubation of oocytes prior to ICSI is associated with improved maturation of oocytes, fertilization and embryo quality.

Adult↗

The combined use of vibrostimulation and in vitro fertilization: successful pregnancy outcome from a retrograde specimen obtained from a spinal cord-injured male.

While pregnancies have been documented through the independent use of the vibrator method, from other methods of procuring ejaculate from spinal cord injured men, and from artificial insemination using a retrograde specimen, we believe that this is the first case report of a live birth resulting from a retrograde ejaculate obtained by vibration from a spinal cord-injured male whose partner underwent in vitro fertilization. Vibrostimulation may well be successful in the two-thirds of men whose spinal cord lesions are at the T10 neurological level and above, who have an intact bulbocavernosus reflex and anal tone but no pain or temperature sensation of the genitalia. Blood pressure monitoring, prevention of autonomic dysreflexia, alkalinization, dilution and infection control of urine, and retrograde specimen retrieval are all important techniques to ensure patient safety and optimal ejaculates. The timing of ovulation and insemination is the crucial factor for the partner of a SCI male whose sperm quality is poor. A complete gynecological workup, including studies of tubal patency, should be done before embarking on a series of artificial inseminations. Stimulation of ovulation and well-timed inseminations should optimize the chance of conception. Depending on semen analysis, female partner factors, and emotional and financial costs, IVF can appropriately be either an early or a final option.

Adult↗

Transfer and uterine factors are the major recipient-related determinants of success with donor eggs.

OBJECTIVE: To define the recipient-related determinants of outcome with donor eggs. DESIGN: Case-control study. SETTING: Community hospital-based assisted reproductive technology (ART) program. PATIENT(S): One hundred thirty-four embryo transfers (ETs) in which two recipients were matched to one donor. INTERVENTION(S): Controlled ovarian hyperstimulation (COH) and ovum retrieval in donors; IVF and ET to recipients. MAIN OUTCOME MEASURE(S): Recipients' age, body mass index (BMI), medical conditions, endometriosis, gravidity, uterine pathology, endometrial thickness, egg number, total motile sperm count, intracytoplasmic sperm injection (ICSI), zygote number, fertilization rate, embryos per ET, embryos frozen, embryo quality, difficulty with transfer, and ongoing pregnancies per ET. RESULT(S): Forty-one recipient pairs had discordant outcomes. Pregnant patients had a lower frequency (9.7% vs. 31.7%, P=.04) and lesser severity of uterine pathology. Endometrium <8 mm was found solely in failed cycles. Pregnant women had fewer moderate or difficult ETs (9.7% vs. 31.7%, P=.04) and more good embryos (1.8 vs. 1.3, P=.03) than the nonpregnant group. CONCLUSION(S): Analysis of recipient pairs with discordant outcomes identifies the recipient-related predictors of success by keeping oocyte quality and the laboratory component constant. Uterine pathology, thin endometrium, transfer difficulty, and number of high-grade embryos are the principal recipient-related determinants of outcome with donor eggs.

Adult↗

Birth of two healthy females after preimplantation genetic diagnosis for familial amyloid polyneuropathy.

Familial amyloid polyneuropathy (FAP), Portuguese type, is a late onset, high penetrance, autosomal dominant Mendelian disorder caused by a V30M substitution in the transthyretin (TTR) protein. A genetic diagnosis was developed using fluorescent single cell polymerase chain reaction (PCR) on lymphocytes from patients and controls. Ovarian stimulation and oocyte retrieval were carried out using conventional protocols in a couple in whom the female was heterozygous for the mutation TTR V30M. Blastomere biopsy was performed on day 3 after intracytoplasmic sperm injection. PCR was then performed for a segment of the TTR gene encompassing the V30M mutation. The transfer of three embryos at day 4 resulted in a twin pregnancy, confirmed as healthy females by amniocentesis at 16 weeks of gestation; the birth took place at 37 weeks of gestation. With this report, FAP, TTR related, joins the lengthening list of genetic conditions for which preimplantation genetic diagnosis has been successfully carried out.

Adult↗

Treatment of retrograde ejaculation and anejaculation.

Treatment of retrograde ejaculation and anejaculation The various options for the treatment of retrograde ejaculation (RE) and anejaculation (AE) are discussed systematically in this review. A total of 88 studies dealing with patients with RE emphasize medical treatment for reversal of RE and retrieval of spermatozoa from urine. In 136 studies concerning patients with AE, the main emphasis is on medical treatment, electroejaculation (EE) and electrovibration stimulation (EVS) for the reversal of AE. Sperm quality in patients with RE and AE is often impaired. However, with the help of assisted reproduction techniques (ART) available today, both ejaculation disorders can be considered as treatable diseases. The major problem when analysing the studies was the uneven methodological quality of the original articles and the difficulties presented by different drugs and dosages, equipment and techniques, along with different criteria for success. In conclusion, controlled clinical trials comparing different treatment options appear urgently warranted.

Ejaculation↗

Comparison of pronuclear stage embryo transfer and in vitro fertilization-embryo transfer following intracytoplasmic sperm injection.

To determine whether the mode of embryo transfer (PROST vs IVF) affected the outcome in intracytoplasmic sperm injection (ICSI) cycles, 237 ICSI cycles (106 PROST and 131 IVF) were analyzed. Several parameters, including patient age, duration of infertility, amounts of hMG used, number of mature eggs retrieved and injected, fertilization rate, number of embryos transferred, and clinical pregnancy rate, were compared. Most of the variable factors were not significantly different, except the mean numbers of transferred embryos which were significantly higher in the PROST group. The clinical pregnancy rate showed no statistical difference between PROST and IVF cycles (25.5 and 16.8%; p = 0.139). This study suggests that even the pregnancy rate in PROST cycles was slightly higher than IVF cycles, but there was no statistically significant difference between the two groups.

Adult↗

Maturation in vitro of human oocytes from unstimulated cycles: selection of the optimal day for ovum retrieval based on follicular size.

The potential use of immature oocytes for in-vitro fertilization (IVF) requires the conditions for successful maturation to be defined. This study focused on the day of oocyte retrieval. The selection of a dominant follicle may induce endocrine changes in the remaining cohort that may be detrimental to their subsequent fertilization and embryonic development. Natural cycles in volunteer donors were followed by measurement of serum oestradiol and by vaginal ultrasound, starting on day 3 of the cycle. Cycles were randomly allocated to one of two groups: group 1 (n = 10), in which follicles were aspirated before the leading follicle was 10 mm in diameter; and group 2 (n = 9), in which follicles were aspirated when a dominant follicle was clearly visible with diameter >10 mm. Oocytes were cultured in vitro to metaphase II (MII) stage, donated, and inseminated by intracytoplasmic sperm injection (ICSI) with husband's spermatozoa. Those that became fertilized within 24 h were further co-cultured in autologous endometrial epithelial cells up to the blastocyst stage, and cryopreserved. There was a significantly (P < 0.05) increased rate of oocyte retrieval in group 1 (70.8% of aspirated follicles) compared with group 2 (50.5%). Maturation to MII and fertilization were similar between the groups. However, development to blastocyst stage was significantly (P < 0.05) higher in group 1 embryos (56.5%) compared with group 2 (35.7%). There was a positive correlation (r2 = 0.1978) between the appearance of the cumulus cells and the ability to develop to blastocyst stage when both parameters were analysed in group 1, whereas no such correlation was found in group 2. In conclusion, our data suggest the importance of retrieving immature oocytes before follicular selection, and define the conditions for the first stage in the use of immature oocytes. Further stages must be defined before this technique can be used clinically.

Adult↗

Fertilization, pregnancy and embryo implantation rates after ICSI with fresh or frozen-thawed testicular spermatozoa.

This study aimed to determine whether fertilization and implantation rates after intracytoplasmic sperm injection (ICSI) with fresh or frozen-thawed testicular spermatozoa were comparable. Between 1 January 1996 and 31 December 1996, 65 ICSI cycles with testicular spermatozoa and 35 cycles with frozen-thawed testicular spermatozoa were carried out. In 50 out of 65 ICSI cycles, testicular spermatozoa could be retrieved and in 34 out of 35 cycles carried out with frozen-thawed testicular spermatozoa, motile spermatozoa could be recovered. The fertilization rate after ICSI with frozen-thawed testicular spermatozoa was significantly lower (71.1%; P < or = 0.008) than with fresh testicular spermatozoa (79.3%). The pregnancy rate was similar for both groups (38.2 and 26.5 %). The implantation rate per transferred embryo, however, was significantly lower in the frozen-thawed rather than in the fresh testicular sperm group (9.1 versus 24.6%; P = 0.001). The live birth rate per transferred embryo was also higher in the group in which fresh testicular spermatozoa were used (18.8 versus 7.9% P = 0.043). This retrospective study shows that is possible to achieve a high fertilization rate after ICSI with both fresh and frozen-thawed testicular spermatozoa but implantation and live birth rates per transferred embryo, however, are significantly lower after ICSI with frozen-thawed than with fresh testicular spermatozoa.

Cryopreservation↗

Outcome of pregnancies derived from assisted reproductive technologies: IVF versus ICSI.

PURPOSE: To compare the course and outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) pregnancies. METHODS: A retrospective study was conducted in a university-affiliated IVF unit with 200 patients who conceived in 1996-1997, 100 with ICSI and 100 with IVF. Data were retrieved from our prospectively created computerized database. In addition, all patients were interviewed by telephone, and the interviewing physician completed a detailed questionnaire. Findings for the IVF and ICSI pregnancies were compared. The main outcome measures were maternal age, implantation rate, early pregnancy complications, clinical abortion rate, multiple pregnancy delivery rate, gestational age at delivery, mode of delivery, and birth weight. RESULTS: In all, 238 children were born, including 104 singleton infants (45 IVF, 59 ICSI), 49 twin pairs (28 IVF, 21 ICSI), and 12 triplet sets (3 IVF, 9 ICSI). Statistically significant differences between the ICSI and IVF groups were noted for maternal age (31.3 +/- 4.4 vs. 33.4 +/- 4.8, respectively, P < 0.005) and clinical abortion rate (11% vs. 24%, respectively, P < 0.05). CONCLUSIONS: ICSI pregnancies in our series were characterized by a lower clinical abortion rate than IVF pregnancies, probably because of the mean younger age of the ICSI group.

Abortion, Spontaneous↗

Application of intracytoplasmic sperm injection in assisted reproductive technologies.

Intracytoplasmic sperm injection (ICSI) entails the mechanical insertion of a chosen spermatozoon directly into the cytoplasm of an oocyte. Due to the consistent fertilization and pregnancy outcome, ICSI is routinely used to treat azoospermic patients where spermatozoa are retrieved by epididymal aspiration or testicular biopsy. Since male subfertility has been associated with a higher incidence of genomic defects, ranging from numerical chromosomal abnormalities to Yq microdeletions, concerns have been raised as to the risk of transmitting genetic defects to the offspring. Screening for such defects can provide invaluable information for appropriate counselling prior to ICSI treatment. In order to address these concerns, a follow-up of the children born after ICSI treatment was conducted.

Adult↗

Generation of normal progeny by intracytoplasmic sperm injection following grafting of testicular tissue from cloned mice that died postnatally.

Animal cloning by nuclear transfer has been successful in several species and was expected to become an alternative reproductive technique. Among the problems associated with this cloning technique, however, are its low success rate and high mortality of cloned animals even if they develop to term. Nuclear transfer has thus come to be considered too difficult to apply as a reproductive technique. The transplantation of male germ cells or pieces of testicular tissue has enabled the induction of spermatogenesis from fetal or postnatal male mice. In the present study, we examined whether functional male gametes could be obtained by the transplantation of pieces of testicular tissue from cloned mice that died immediately after birth with typical aberrant phenotypes, such as large offspring syndrome. Donor testicular tissues were retrieved from cloned mice that died postnatally and were transplanted into the testes of recipient nude mice. Two to three months after transplantation, the grafted donor testicular tissue had grown in the host testis, and histological analysis showed that spermatogenesis occurred within the graft. Intracytoplasmic sperm injection demonstrated that the testicular sperm generated in the grafted donor tissue were able to support full-term development of progeny. These results clearly showed that functional spermatogenesis could be induced by transplanting testicular tissue from cloned mice that died postnatally into recipient mice. The strategy presented here will be applicable to cloned animals of other species, because the xenografting of testicular tissue into mice has been demonstrated previously to be possible.

Animals↗

Combined diagnostic laparoscopy and follicular aspiration for human in vitro fertilization.

Laparoscopic retrieval of oocytes from stimulated follicles for in vitro fertilization and embryo transfer (IVF-ET) was performed at the time of diagnostic laparoscopy in 36 infertile women. Their mean rates of cleavage, embryo transfer and pregnancy did not differ from those obtained in 86 patients (101 cycles) who underwent ultrasonically guided transvaginal oocyte retrieval following diagnostic laparoscopy performed earlier as a separate procedure. In view of the advantages in terms of reduced patient morbidity, time and cost benefits, and the opportunity for oocyte-sperm compatibility testing in vitro, the combined procedure appears to be the best method in patients scheduled to undergo diagnostic laparoscopy.

Adult↗

Developmental ability of human oocytes with or without birefringent spindles imaged by Polscope before insemination.

BACKGROUND: Birefringent spindles imaged with the Polscope can predict fertilization rates after intracytoplasmic sperm injection (ICSI). The present study examined the development of human oocytes with or without birefringent spindles, imaged with the Polscope before ICSI. METHODS: Oocytes were obtained from stimulated ovaries of consenting patients undergoing oocyte retrieval for ICSI. Spindles were imaged with the Polscope combined with a computerized image analysis system. After imaging and ICSI, oocytes with or without spindles were cultured separately for examination of fertilization and embryo development. A total of 1544 oocytes from 136 cycles were examined with the Polscope and inseminated by ICSI. RESULTS: Spindles were imaged in 82% of oocytes. After ICSI, more oocytes (P < 0.05) with spindles (69.4%) fertilized normally, forming 2 pronuclei, than oocytes without spindles (62.9%). At day 3, more oocytes (P < 0.01) with spindles (66.3%) developed to 4-11 cell stages than oocytes without spindles (55.4%). Significantly more (P < 0.001) oocytes with spindles developed to morula and blastocyst by day 5 (51.1 versus 30.3%) and day 6 (53.2 versus 29.3%) compared with oocytes without spindles. CONCLUSIONS: The results indicate that the presence of a birefringent spindle in human oocytes can predict not only higher fertilization rate, but also higher embryo developmental competence.

Adult↗

Controlled comparison of conventional in-vitro fertilization and intracytoplasmic sperm injection in patients with asthenozoospermia.

A controlled comparison between conventional in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) has been carried out for patients with </=5% rapid progressive (type A) spermatozoa in their fresh semen. Twenty couples were allocated to the study. All semen samples fulfilled the minimum criteria to be allocated to the conventional IVF programme. The two pronuclear (2PN) fertilization rate per retrieved cumulus-oocyte complex was significantly higher after ICSI (63.4%) than after conventional IVF (22.9%), with complete fertilization failure after IVF in 10 out of the 20 cycles. Embryo quality was similar for both treatments. Sixteen patients received only ICSI embryos, two patients only IVF embryos and two patients received IVF and ICSI embryos. The ongoing clinical pregnancy rate was 45%, the implantation rate was 37%. Comparison of the characteristics of patients/cycles with and without 2PN fertilization revealed a higher proportion of progressively motile spermatozoa in the prepared sperm fraction for the group of patients with fertilization after conventional IVF. It can be concluded from this study that absence of or an extremely low proportion of rapid progressive motility in fresh semen indicates a high risk of complete fertilization failure with conventional IVF.

Embryo Implantation↗

Sperm morphology and IVF: embryo quality in relation to sperm morphology following the WHO and Krüger's strict criteria.

BACKGROUND: The aim of this study was to examine the correlation between sperm morphology and embryo quality/IVF outcome. METHODS: The implication of sperm morphology assessment before an IVF cycle was evaluated. A total of 100 IVF couples where the female partner had either tubal factor (n=50) or unexplained infertility (n= 50) entered a prospective study, and sperm samples for the actual cycle were assessed according to the strict criteria and WHO criteria. The study was blinded for the technician involved in sperm morphology analyzing. IVF was carried out according to a long down regulation protocol using GnRH/FSH/hCG and ova were inseminated with 200,000 spermatozoa/ml. Embryos were transferred on day 2 post fertilization in a maximum of three embryos. RESULTS: No significant differences were found between the groups regarding age of the female partner (mean=34.3), no. oocytes retrieved (mean=8.5), fertilization (66.5%), pregnancies (pos. S-hCG/transfer 39.6%) or 'Take home baby rate' (birth rate/transfer 30.0%). As to the score of Krüger's strict criteria and the WHO criteria, we found no correlation between this score and cleavage rate, embryo development or pregnancies. The WHO criteria were found to be a better predictor for fertilization rate than the Krüger's criteria (p<0.002). CONCLUSION: The strict criteria or sperm evaluation according to WHO have no better predictive value for the outcome of routine IVF.

Adult↗

Medical and psychological management of recurrent abortion, history of postneonatal death, ectopic pregnancy and infertility: successful implementation of IVF for multifactorial reproductive dysfunction. A case report.

The medical and psychological treatment for a 37-year-old Caucasian G6 P1051 woman who presented for evaluation of secondary infertility and recurrent pregnancy loss is described. Although one living child had been conceived without medical assistance, that delivery preceded the present evaluation by ten years and involved a different partner. With the current husband, the patient had two miscarriages and a left ectopic pregnancy. The couple had attempted controlled ovarian hyperstimulation and in vitro fertilization (IVF) elsewhere, but the cycle was cancelled due to poor follicular response. About one year before consultation at our institution, the couple established a pregnancy although the infant was born at 24 weeks with a cardiac anomaly, living only 40 days. Additionally, a persistent cervical lesion required cone biopsy before any fertility treatment could resume. Andrology evaluation found the husband's sperm DNA fragmentation index to be 48.6%. This constellation of stressors represented substantial emotional issues and psychological therapy/counseling was recommended. After obtaining psychological clearance, the couple underwent IVF and 16 oocytes were retrieved. Four embryos were transferred, and a healthy male infant was delivered at term. Although multifactorial infertility can be associated with very poor reproductive outcomes, the advanced reproductive technologies merit consideration during management of complex clinical challenges. Standard IVF strategies can be optimized by inclusion of thorough psychological assessment and counseling.

Abortion, Habitual↗