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PubMed1999

Pregnancies resulting from in vitro matured oocytes retrieved from patients with polycystic ovary syndrome after priming with human chorionic gonadotropin.

OBJECTIVE: To describe pregnancies that resulted from in vitro matured oocytes derived from two unstimulated, anovulatory patients with polycystic ovary syndrome after priming with hCG before oocyte retrieval. DESIGN: Case series. SETTING: McGill Reproductive Center, Royal Victoria Hospital, McGill University. PATIENT(S): Two women with polycystic ovary syndrome. INTERVENTION(S): Progesterone induction of a withdrawal bleeding episode. The administration of subcutaneous hCG 36 hours before oocyte retrieval in a natural menstrual cycle. In vitro maturation of immature oocytes, fertilization, and ET. Luteal support with oral estrogen and progesterone. MAIN OUTCOME MEASURE(S): Pregnancy outcome. RESULT(S): Two clinical pregnancies were confirmed after oocyte maturation in vitro, fertilization with intracytoplasmic sperm injection, and ET. CONCLUSION(S): The administration of hCG 36 hours before harvesting of immature oocytes may improve the maturational and developmental competence of the oocytes and the pregnancy rates of unstimulated patients with polycystic ovary syndrome.

Adult↗
PubMed1999

In-vitro maturation of round spermatids using co-culture on Vero cells.

In an attempt to determine whether co-culture could promote sperm maturation, three patients with non-obstructive azoospermia, two with maturation arrest at the level of primary spermatocytes and one patient with <1% tubules showing complete spermatogenesis, and one patient with total globozoospermia, gave consent to experimentally co-culture round spermatids retrieved from the testicle on Vero cell monolayers. In all azoospermic patients elongating spermatids could be obtained from round spermatids. In one case of maturation arrest, of 37 round spermatids co-cultured for up to 5 days, 30% developed flagella, 46% matured to elongating and 19% to elongated spermatids, with one mature spermatozoon also obtained (3%). In the same patient, primary cultures of three round spermatids with flagella enabled development of one further mature spermatozoon. In the case with total globozoospermia, of six round spermatids co-cultured for up to 5 days, one mature spermatozoon was obtained, with a flagellum and normal head morphology. These preliminary findings suggest that it may be possible to overcome the round spermatid block, and even the triggering of morphological abnormalities arising at the spermiogenic level, by in-vitro maturation under special environmental conditions.

Animals↗
PubMed2001

Ultrastructural evaluation of recurrent and in-vitro maturation resistant metaphase I arrested oocytes.

An infertile couple whose female partner showed recurrent retrieval of immature metaphase I (MI) oocytes that were resistant to in-vitro maturation, was studied. Four spermiograms revealed teratozoospermia. Consistent non-fertilization and negative pregnancy outcomes were obtained after intrauterine insemination, gamete intra-Fallopian transfer and IVF. Two intracytoplasmic sperm injection (ICSI) cycles were finally performed. All oocytes (n = 17) in both cycles were arrested at MI and failed to mature after 48 h culture. ICSI also resulted in total non-fertilization. In the last cycle, two oocytes were analysed by transmission electron microscopy and showed almost identical results. All organelles showed normal characteristics of an MI oocyte. The main abnormality found was related to the MI spindle, with absence of microtubules and dispersion of the female chromosomes. Minor abnormalities were observed (immature fibrous appearance of the zona pellucida; the presence of small vesicle aggregates which formed a foam-like body). The injected sperm nucleus was arrested in the middle of the chromatin decondensation process, with no visible nuclear envelope reformation. Normal disruption of sperm acrosomal and flagellar components were observed. Only a partial cortical reaction was observed. This represents the first documented case of developmental arrest due to complete absence of spindle formation in association with an otherwise mature ooplasm.

Acrosome↗
PubMed2003

[Successful preimplantation genetic diagnosis for beta-thalassemia using primer extension preamplification].

OBJECTIVE: To achieve preimplantation genetic diagnosis (PGD) of the couples at risk of having children with beta-thalassemia, as an alternative to prenatal diagnosis. METHODS: Two couples carrying different thalassemia mutations of codon 41/42 and codon intervening sequence 2 position 654 received standard in vitro fertilization treatment and intracytoplasmic sperm injection, embryo biopsy and the whole genome was amplified by primer extension preamplification (PEP). Nested polymerase chain reaction was then used to amplify two mutation sites separately. Both were detected by reverse dot-blot. RESULTS: A total of 35 oocytes were retrieved from the two patients. Among them, 87% showed two pronuclei, and embryo biopsy was performed on 16 of these embryos and 25 blastomeres were obtained. The amplification efficacy was 84%. The genotype study of non-transferred and surplus embryos showed 15% of allele drop-out rate. Five embryos were transferred to the uterus of both patients. One pregnancy achieved, resulted in live healthy twin births, which confirmed the results of PGD. CONCLUSIONS: This unaffected pregnancy resulting from PGD by PEP for beta-thalassemia demonstrates that this technique can be a effective diagnostic tool for carrier couples who desire a healthy child.

Adult↗
PubMed2001

Human menopausal gonadotropin versus recombinant follicle-stimulating hormone in normogonadotropic women down-regulated with a gonadotropin-releasing hormone agonist who were undergoing in vitro fertilization and intracytoplasmic sperm injection: a prospective randomized study.

OBJECTIVE: To evaluate clinical and endocrinological effects of intranasal (IN) vs. subcutaneous (SC) GnRH-a for pituitary down-regulation combined with hMG vs. rFSH. DESIGN: Prospective, randomized study. SETTING: University hospital, IVF unit. PATIENT(S): Three hundred seventy-nine normogonadotropic women eligible for IVF or ICSI. INTERVENTION(S): Randomization to intranasal (IN) or SC GnRH-a and to hMG or rFSH. MAIN OUTCOME MEASURE(S): Oocytes retrieved, embryos developed, clinical pregnancy, and delivery rates. Serum hormone concentrations on stimulation days 1 (S1) and 8 (S8), and oocyte pick-up (OPU) day. RESULT(S): After randomization, four groups were formed: IN/hMG (n = 100), IN/FSH (n = 98), SC/hMG (n = 89), and SC/FSH (n = 92). Mean number of oocytes retrieved and of transferable and transferred embryos were similar in the four groups. Clinical pregnancy rate per started cycle was significantly higher in the IN/HMG group than in the SC/FSH group (P<.05) and was intermediate in the two remaining groups. Se-LH on S8 in the two SC groups was significantly lower than in the two IN groups. Se-E2 on S8 in the SC/FSH group was significantly lower than in the other three groups. CONCLUSION(S): The clinical and endocrinological outcome in IVF and ICSI-treated normogonadotropic women is significantly influenced by mode of down-regulation as well as gonadotropin formulation.

Administration, Intranasal↗
PubMed2001

Effects of growth hormone on in vitro maturation of germinal vesicle of human oocytes retrieved from small antral follicles.

PURPOSE: Evaluation of the effect of GH, administered in vivo, on in vitro maturation of human GV oocytes obtained from small follicles. METHODS: The study included 88 ICSI women stimulated with long agonist protocol. They were randomly allocated to two groups (Group 1, 44 women with 86 GV oocytes), were cotreated with GH (Group 2, no GH: 44 women with 67 GV oocytes). GV oocytes from small follicles were matured in vitro, injected, and their development observed. RESULTS: Resumption of meiosis was significantly higher in Group 1 compared to Group 2 (60 and 37.3% respectively). Significantly more M2 oocytes were obtained in Group 1 than in Group 2 (45.5 and 23.9% respectively). Fertilization rates were also significantly higher (51.9 and 18.8% respectively). No embryos were obtained in Group 2, whereas 21.1% of fertilized oocytes in Group 1 cleaved into class A embryos. CONCLUSION: In vivo administration of GH enhanced in vitro maturation and fertilization of human GV oocytes retrieved from small antral follicles.

Adult↗
PubMed1999

Prognostic factors in patients continuing in vitro fertilization or intracytoplasmic sperm injection treatment and dropouts.

OBJECTIVE: To investigate whether cumulative pregnancy rates using life table analysis but without considering dropouts are representative of the whole population of patients entering an in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) program. DESIGN: Retrospective study. SETTING: University hospital-based infertility center. PATIENT(S): One thousand one hundred sixty-nine patients entering our IVF/ICSI program from January 1993 to December 1996. INTERVENTION(S): Comparison of prognostic factors between pregnant and nonpregnant patients, and between patients continuing IVF/ICSI treatment and dropouts. MAIN OUTCOME MEASURE(S): Prognostic factors, such as patient age, cancellation of oocyte retrieval because of poor response to ovarian stimulation, number of oocytes retrieved, fertilization rate, number and quality of embryos transferred. RESULT(S): No statistical differences in prognostic factors were found between patients continuing IVF/ICSI treatment and dropouts. CONCLUSION(S): Cumulative pregnancy rates using life table analysis can be considered representative of the whole population of patients for at least the first three treatment cycles.

Adult↗
PubMed1997

Transport-in-vitro fertilization/intracellular sperm injection: a prospective randomized study.

We performed a prospective randomized clinical trial to investigate whether long distance oocyte transport prior to an intracytoplasmic sperm injection (ICSI) procedure influences fertilization rates, embryo quality and/or embryo implantation rates. After informed consent, 100 infertile couples booked for ICSI treatment were randomized into two groups. In group 1 (n = 50), patients were stimulated and monitored in Rotterdam (The Netherlands), and oocyte retrieval, ICSI procedure and embryo transfer took place in Gent (Belgium). In group 2 (n = 50), patients were stimulated, monitored and punctured in Rotterdam and the oocytes were transported in their follicular fluid in an isothermic transport box to Gent, where the ICSI procedure and the embryo transfer took place. In both groups the stimulation and monitoring regimen, puncture technique, laboratory conditions and transfer policy were identical. In both groups, the number of fertilized oocytes (7.13 +/- 0.65 versus 5.53 +/- 0.60, P = 0.08), the number of transferred embryos (2.36 +/- 0.09 versus 2.40 +/- 0.11, P = 0.87) and the embryo implantation rate [20/113 (17.7%) versus 19/103 (18.4%), P = 0.89] was similar. In group 1, the number of retrieved oocytes was higher (10.83 +/- 0.95 versus 8.44 +/- 0.93, P = 0.05). The total score of the embryos obtained (18.90 +/- 1.73 versus 12.64 +/- 1.26, P = 0.01), the number of good quality embryos (4.63 +/- 0.49 versus 2.98 +/- 0.38, P = 0.02), the mean score of the transferred embryos (3.32 +/- 0.11 versus 2.94 +/- 0.13, P = 0.05) and the number of embryos available for cryopreservation (2.70 +/- 0.45 versus 1.48 +/- 0.38, P = 0.03) were significantly higher in group 1. Therefore, long distance transport of oocytes prior to ICSI does not affect oocyte fertilization and embryo implantation rates, although a negative effect on embryo quality cannot be excluded.

Adult↗
PubMed1999

Pregnancies after activated oocyte transfer: a new option for infertility treatment.

In this preliminary report, we describe a new technique involving the same-day transfer of activated oocytes to the uterus after intracytoplasmatic sperm injection (ICSI). The technique, termed activated oocyte transfer (AOT), offered to 19 couples, yielded a pregnancy rate per cycle of about 30%, equivalent to traditional in-vitro fertilization (IVF) and ICSI in a laboratory setting. AOT is performed 4 h after oocyte retrieval, permitting the patient to undergo treatment as an out-patient procedure.

Adult↗
PubMed2003

The serum concentration of estradiol after embryo transfer and the decline from preovulatory levels may influence the success of IVF treatment.

AIM: To establish the influence that serum estradiol concentrations prior to oocyte retrieval and 3 days after embryo transfer have on the establishment of in vitro fertilization (IVF) pregnancy. METHOD: Preovulatory (day-0) and luteal-phase (day-6) estradiol concentrations were retrospectively analyzed in 310 infertile patients, undertaking 1st-cycle conventional IVF or intracytoplasmic sperm injection. RESULTS: The IVF treatment success is significantly reduced in patients with an estradiol level <600 pg/ml and also when a rapid decline in luteal-phase estradiol concentrations from preovulatory concentrations (day 0:day 6 ratio) was exhibited. CONCLUSION: A day 0:day 6 estradiol ratio >5 and a serum estradiol concentration <600 pg/ml may adversely impact on the establishment of pregnancy in IVF.

Adult↗
PubMed2002

Rigorous thermal control during intracytoplasmic sperm injection stabilizes the meiotic spindle and improves fertilization and pregnancy rates.

OBJECTIVE: To examine the effects of different thermodynamic control systems on the temperature stability of human eggs during in vitro manipulation, with the integrity of meiotic spindles imaged using the LC-PolScope (Cambridge Research & Instrumentation, Inc., Woburn, MA). DESIGN: We performed intracytoplasmic sperm injection (ICSI) and/or imaging of eggs with the temperature regulated by three different systems: thermostated coverslip (system 1), thermostated coverslip combined with objective heater (system 2), and conventional stage warmer (system 3). SETTING: Academic in vitro fertilization clinic. PATIENT(S): Oocytes were aspirated from stimulated ovaries of patients undergoing oocyte retrieval for ICSI. INTERVENTION(S): Measurement of temperature regulation in media surrounding eggs during in vitro manipulation and imaging. MAIN OUTCOME MEASURE(S): Rate of oocytes with spindles, fertilization rates, and clinical pregnancy rates after ICSI. RESULT(S): We imaged spindles in more oocytes with system 2 (81.2%) than with system 1 (61.4%). Spindles could not be imaged for system 3 because of technical limitations. Fertilization rates were significantly higher when oocytes were imaged and used for ICSI with system 2 (78.8%) than with system 1 (56.7%) or system 3 (64.0%). Most importantly, a significantly higher clinical pregnancy rate was observed when oocytes were manipulated with system 2 (51.7%) than with system 1 (25.0%) or system 3 (23.1%). No differences were found in average ages, number of previous cycles, number of eggs, or day 3 FSH or E2 levels among groups. CONCLUSION(S): Imaging meiotic spindles with the PolScope provides an intracellular thermostat during ICSI. Rigorous thermal control during ICSI stabilized spindles and increased the fertilization and clinical pregnancy rates achieved after ICSI. The presence of birefringent spindles in living human eggs served as a monitor for in vitro conditions.

Adult↗
PubMed2000

Influence of female age on pregnancy outcome in in vitro fertilization and embryo transfer patients undergoing intracytoplasmic sperm injection.

PURPOSE: To determine the influence of female age on the outcomes of ICSI in IVF-ET patients. METHODS: One hundred and seventy-five couples underwent 352 cycles of ICSI. The quality of oocytes and embryos, fertilization rate, and pregnancy outcomes were retrospectively evaluated according to female age; < 30 years in Group A (49 cycles), 30-34 in Group B (177 cycles), 35-39 in Group C (97 cycles), and > or = 40 in Group D (29 cycles). RESULTS: The fertilization rates were not significantly different among the age groups. Significant negative linear correlations were observed between female age and the numbers of oocytes retrieved and embryos transferred, and cumulative embryo score. Clinical pregnancy rates were significantly decreased and spontaneous abortion rate increased with advancing age. CONCLUSIONS: Female age may be a prognostic indicator in ICSI program.

Adult↗
PubMed1989

Improvement of survival and fertilizing capacity of human spermatozoa in an IVF programme by selection on discontinuous Percoll gradients.

Two techniques for the separation of spermatozoa were compared: swim-up migration (SUM) and centrifugation on a discontinuous Percoll gradient (CPG). Their respective effects on sperm motility were analysed by computer-assisted videomicrography in either normal or asthenozoospermic groups. In both groups, there was no difference in any of the motion parameters between the two treatments after 1-h incubation. However, a clear difference was observed after 24 h when excellent motility was retained only in the CPG-treated group. A total of 350 ejaculates were produced by the husbands of women undergoing oocyte retrieval in an IVF programme. Spermatozoa were treated by CPG when the infertility was due to poor quality spermatozoa (n = 91), when there was a known previous history of semen infection (n = 73) or when frozen semen, originating from a donor, was used (n = 36). In all other cases (n = 150), spermatozoa were treated by SUM. The cleavage rates obtained were 32.2, 70.1, 60.9 and 68.6% respectively in the four categories. The clinical pregnancy rates per oocyte retrieval were 19.8, 31.5, 22.2 and 18.0% respectively. Forty-eight births occurred in the CPG group: 28 boys and 20 girls, all normal. We conclude that CPG is useful, both in cases of poor semen quality and in tubal infertility, in which the clinical pregnancy rate increased significantly from 18.0 to 31.5%.

Cell Survival↗
PubMed2004

Semen characteristics in human immunodeficiency virus (HIV)- and hepatitis C (HCV)-seropositive males: predictors of the success of viral removal after sperm washing.

BACKGROUND: Human immunodeficiency virus (HIV)/hepatitis C virus (HCV)-seropositive males can now father children safely, avoiding transmission risks to the mother and the children using sperm washing and nested PCR (nPCR) techniques. Nevertheless, we still lack enough data to determine the reasons why approximately 10% of the performed sperm washes remain positive, thus forcing the repetition of the treatment. Semen quality in infected males is also essential to these procedures. We aimed to determine the predictive value of the semen parameters, sperm washing procedure and the infection status for the post-wash viral positivity, as well as the correlation between the semen and the disease features. METHODS: Semen characteristics were evaluated in 136 samples provided from 125 males. We also included a control group of 125 males matched by age and length of sexual abstinence. At the time of semen retrieval, 70 of them were infected with HIV (45 also with HCV, 64.3%), and 55 of them with HCV alone. nPCR for viral detection was performed in each sample. RESULTS: Thirteen out of 136 (9.5%) of the samples were positive for one or more viral detections (HIV RNA, HIV DNA and HCV RNA, when needed). From a total of 240 nPCR viral analyses, 16 were positive (6.6%). None of the seminal parameters were adequate to predict post-wash results, nor was a positive result dependent on the volumes used in the semen wash. A positive correlation was found between post-wash progressive motility and CD4 blood levels, as well as a negative correlation between progressive motility and time of evolution of the disease in HIV-infected males. CONCLUSIONS: Semen analysis, according to the World Health Organization criteria, of HIV- and HCV-affected patients showed no differences from that of non-infected males. Moreover, low CD4 blood levels, and a long evolution of the disease do not negatively affect sperm motility.

Anti-Retroviral Agents↗
PubMed2006

Laser versus mechanical assisted hatching: a prospective study of clinical outcomes.

OBJECTIVE: To compare clinical outcomes after laser or mechanical techniques of assisted hatching (AH) in women of advanced age undergoing IVF or intracytoplasmic sperm injection (ICSI). DESIGN: Prospective randomized study. SETTING: Center for Human Reproduction, Genesis Clinic, Athens, Greece. PATIENTS: Three hundred and sixteen consenting women of advanced age (> or =39 years) with primary infertility undergoing IVF/ICSI programs and having available embryos for transfer on day 3. Patients were randomized into laser or mechanical AH of their transferred embryos. INTERVENTIONS: Controlled ovarian hyperstimulation, oocyte retrieval, IVF/ICSI, laser or mechanical AH, and embryo-transfer. MAIN OUTCOME MEASURES: Implantation rate, clinical pregnancy rate, and viable pregnancy rate. RESULTS: The implantation rate was significantly higher in the laser AH group. Clinical and viable pregnancy rates were higher (but not significantly) in the laser AH group. CONCLUSIONS: Laser AH of embryos may result in better clinical outcomes when compared to the mechanical technique in women of advanced age undergoing IVF or ICSI.

Adult↗
PubMed2006

Timed IVM followed by ICSI in a patient with immature ovarian oocytes.

A complete failure of meiotic maturation occasionally occurs following human chorionic gonadotrophin administration during IVF-intracytoplasmic sperm injection (ICSI) cycles. ICSI on day 1 is commonly used to allow maturation in culture. However, if the oocytes become mature in the evening soon after their recovery but ICSI is delayed until the next day, then subsequent ageing of matured oocytes may be unfavourable for fertilization and development. To avoid the deterioration associated with oocyte ageing, the timing of polar body extrusion was checked every 3 h and rescue in-vitro maturation (IVM)-ICSI was performed shortly after the polar body extrusion was confirmed. This report describes a successful pregnancy and birth of a healthy baby in a patient who had no mature oocytes at the time of oocyte retrieval, and illustrates the value of extra monitoring for IVM and ICSI in cases where only immature oocytes are available.

Adult↗
PubMed1999

A comprehensive approach to the management of male infertility following spinal cord injury.

OBJECTIVES: To review the outcomes of management of male infertility following spinal cord injury in a specialised fertility clinic. STUDY DESIGN: Retrospective review of medical records. SETTING: The Fertility Clinic of a 45-bed spinal cord injury service in New South Wales, Australia. METHODS: The medical and fertility clinic records of 113 males who attended the clinic between 1987 and 1997 were reviewed. RESULTS: The rates of semen retrieval using vibroejaculation and electroejaculation were 67% and 97% respectively. Thirty-one individuals (37%) and their partners sought assistance with the primary aim of achieving a pregnancy. Intravaginal insemination (IVI) undertaken at home following vibroejaculation resulted in a pregnancy rate per cycle of 22%. Electroejaculation and IVI proved less successful with a pregnancy rate per cycle of only 5%, although with intrauterine insemination (IUI) this rate improved to 30%. Micromanipulation in vitro fertilisation (IVF) procedures, primarily intracytoplasmic sperm injection, were used in 18 couples resulting in a pregnancy rate per cycle of 19%. In the 31 couples there have been a total of 17 pregnancies in 97 cycle attempts for an overall pregnancy rate per cycle of 18% and a cumulative pregnancy rate per couple of 55%. Twelve of the pregnancies have resulted in 14 live births (including two sets of twins), there were three pregnancies ongoing at the date of review and there have been two spontaneous abortions. CONCLUSION: The benefits of a specialised fertility clinic offering a comprehensive, client-focused approach with education, fertility assessment and a range of semen retrieval and assisted reproduction options, are highlighted.

Adolescent↗
PubMed2004

Triptorelin versus cetrorelix in intracytoplasmic sperm injection cycles in women with a single ovary.

OBJECTIVE: To compare the response to ovarian stimulation with either the long protocol of a GnRH-agonist or the multiple protocol of a GnRH-antagonist, in women with a single ovary who underwent intracytoplasmic sperm injection (ICSI) cycles. STUDY DESIGN: Retrospective study including 75 ICSI cycles from 26 women. Sixty-three cycles were stimulated with triptorelin/hMG or rFSH, whereas 12 cycles were stimulated with cetrorelix/hMG or rFSH. RESULTS: There was not found any statistical significant difference between the two groups regarding the days of stimulation, the number of gonadotropins' ampoules, the peak estradiol levels, the number of aspirated follicles and the number of retrieved oocytes. The fertilization rate, the number of transferred embryos as well as the cumulative embryo score were also similar in both groups. CONCLUSION: The multiple stimulation protocol of cetrorelix is equally effective with the long protocol of triptorelin in the ovarian stimulation of women with a single ovary.

Adult↗