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PubMed1995

Cryopreservation of human oocytes and fertilization by two techniques: in-vitro fertilization and intracytoplasmic sperm injection.

Human oocyte cryopreservation results in poor survival and subsequent fertilization rates. It has been suggested that freeze-thaw-induced changes in the zona pellucida may impair sperm penetration or attachment. The aim of this study was to compare fertilization and cleavage rates in cryopreserved oocytes inseminated by conventional in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). A total of 220 oocytes, obtained from volunteers who had undergone ovarian stimulation, were cryopreserved using a slow freeze-rapid thaw protocol with 1.5 M propanediol as the cryoprotectant. Surviving oocytes (n = 74, 34.4%) were randomly allocated for fertilization by conventional IVF (group 1) or ICSI (group 2) using cryopreserved spermatozoa from a single donor of proven fertility. Fertilization was achieved in five (13.5%) of the oocytes in group 1 and 17 (45.9%) in group 2 (P < 0.005), with only one oocyte in group 1 exhibiting normal fertilization as opposed to 16 (43.2%) in group 2 (P < 0.001). Similarly, one oocyte fertilized by IVF cleaved, while all fertilized with ICSI cleaved (P < 0.001). We conclude that although the survival of oocytes is poor following cryopreservation, fertilization and cleavage rates can be enhanced significantly using ICSI. These data also suggest that the method of cryopreservation used in this study affected the zona pellucida, such that normal sperm attachment or penetration was impaired.

Cleavage Stage, Ovum↗
PubMed1989

Enhanced viability after in vitro fertilization of bovine oocytes matured in vitro with high concentrations of luteinizing hormone.

The purpose of this study was to better understand requirements for oocyte maturation to yield viable embryos after bovine in vitro fertilization (IVF). High proportions (95% to 100%) of cumulus-surrounded oocytes matured in vitro in all treatments, but subsequent development was enhanced after maturation with high concentrations of purified bovine luteinizing hormone (LH). Beneficial effects of undisturbed cumulus cells were demonstrated. Improved IVF followed insemination of cumulus-surrounded oocytes, but not denuded oocytes, after maturation with high LH (100 micrograms/ml) versus low LH (10 micrograms/ml), implicating cumulus cells in mediating hormonal enhancement. Oocytes matured with high LH resulted in embryos of superior viability, as reflected by cleavage to 4- to 8-cell stages. Pregnancy resulting from transcervical embryo transfer further documented embryonic viability. These findings should be useful in further development and implementation of reproductive and genetic technologies.

Animals↗
PubMedDate not supplied

Correlation between human sperm swelling in hypoosmotic medium (hypoosmotic swelling test) and in vitro fertilization.

Human ejaculates (n = 83) were analyzed for standard sperm parameters (concentration, motility, and morphology), as well as for the ability of the spermatozoa to react (swell) in a hypoosmotic medium (Jeyendran et al, 1984). Subsequently, the fertilizing capacity of the spermatozoa was tested by their ability to fertilize human oocytes in vitro. Although the sperm concentration was adjusted for in vitro fertilization, no adjustments were made for sperm motility and morphology. Correlation of the in vitro fertilizing capacity of the spermatozoa with the hypoosmotic swelling test (r = 0.56) was much higher than with standard sperm parameters (r varied from -0.04 to 0.25). Complete overlap was noted with standard semen parameters whether the ejaculate did or did not fertilize oocytes and ranged from very low to very high values in both cases. By contrast, all the semen samples that fertilized oocytes showed a 60% or higher reaction in the hypoosmotic swelling test, whereas the majority of the "infertile" semen samples showed less than 60% swelling. It therefore appears that, under the conditions of our studies, the hypoosmotic swelling test is a more accurate predictor of successful in vitro fertilization outcome than the conventional semen parameters. A combination of all parameters, however, is likely to be most useful. The hypoosmotic swelling test is simple and economical, and it is recommended that this test be further scrutinized for its value as an additional tool in the assessment of the in vivo fertilizing capacity of ejaculated spermatozoa.

Cell Survival↗
PubMed1994

An indiscriminate use of pentoxifylline does not improve in-vitro fertilization in poor fertilizers.

In order to enhance fertilization in vitro, pentoxifylline (PTX) was used in couples showing low fertilization rates in previous in-vitro fertilization (IVF) attempts for the treatment of male-factor infertility. Sibling oocytes were inseminated at random with spermatozoa prepared with or without PTX. After selection by Percoll gradient, sperm samples were divided into two equal aliquots. One aliquot was incubated in Earle's medium containing approximately 3.6 mM PTX (treatment group), the other aliquot was incubated with PTX-free Earle's medium (control group). After 30 min, both suspensions were washed twice. Sperm parameters after preparation did not differ between treatment and control samples, and nor did the mean fertilization rates, which were 49.3 and 42.6% respectively. Cleavage characteristics and morphological quality of the embryos were not significantly different between the treatment and control groups. The results of this study demonstrate that indiscriminate use of PTX in an IVF programme increases neither fertilization rate nor embryo transfer rate in poor fertilizers. More prospective research is needed to evaluate the role of PTX in IVF in couples selected according to the effect of the compound on sperm function, e.g. hyperactivation and acrosome reaction.

Adult↗
PubMed2004

Use of the egg-share model to investigate the paternal influence on fertilization and embryo development after in vitro fertilization and intracytoplasmic sperm injection.

OBJECTIVE: To investigate whether sperm from different males can influence fertilization and embryo development. DESIGN: To use an egg-sharing model, in which the eggs from one woman are shared between herself and a recipient, and different spermatozoa are used to fertilize the eggs. SETTING: Assisted Conception Unit, Birmingham Women's Hospital, Edgbaston, United Kingdom. PATIENT(S): Infertile women undergoing egg sharing. INTERVENTION(S): In vitro fertilization (IVF). MAIN OUTCOME MEASURE(S): Fertilization rates and the mean day 2 or 3 embryo score (cell number X grade) were examined for egg-sharing pairs. A comparison was also made for pairs in which intracytoplasmic sperm injection (ICSI) and IVF was used as the insemination method. A paired samples t-test was used to compare the sharer and recipient results. RESULT(S): Pregnancy rates did not differ between sharer and recipient couples. Interestingly, when comparing fertilization, there was a significant difference (P<.05) in favor of IVF over ICSI. When comparing embryo development between egg-sharing pairs, we found that approximately 30% of patients showed a difference in mean embryo score of >or= 5 in all embryo development and 14% in the quality of embryos available for transfer. CONCLUSION(S): We showed that the egg-sharing model is a successful alternative for the treatment of women who required donated eggs. More important, the egg-sharing model shows that, in a certain percentage of couples, differences in early embryo development are paternally influenced.

Embryo, Mammalian↗
PubMed1988

Failed fertilization during an in vitro fertilization cycle after oral ingestion of amantadine hydrochloride.

Oocyte fertilization occurred during three in vitro fertilization/embryo transfer (IVF/ET) treatment cycles of an infertile couple but failed during an IVF cycle when the husband took amantadine for prophylaxis against viral infection. Semen parameters were similar to those of other cycles attempted by this couple as well as to those of other IVF couples treated concurrently. We circumstantially suggest that amantadine, and potentially other ingestible medications or foods, while not spermicidal, may impair gamete function during IVF/ET.

Adult↗
PubMed1999

Sperm treatment with extracellular ATP increases fertilization rates in in-vitro fertilization for male factor infertility.

Previous work from our laboratory has revealed that extracellular ATP is a rapid and potent activator of human sperm acrosome reaction and fertilizing ability. In the present study, we assessed the effects of in-vitro sperm incubation with ATP on fertilization and embryo development in couples undergoing in-vitro fertilization (IVF) for male factor infertility. Oocytes from 22 women undergoing ovulation induction were divided in two groups and inseminated in vitro either with selected spermatozoa from the corresponding partner suffering from male factor infertility pre-incubated with ATP (2.5 mM) for 1 h, or with spermatozoa incubated with 0.9% NaCl solution (control group). After insemination, fertilization was assessed by the presence of pronuclei and then by embryo cleavage. The fertilization rate in the group of oocytes inseminated with ATP-treated spermatozoa improved significantly with respect to the control group (65.7 versus 42.5%, P < 0.01). No significant differences were observed in embryo cleavage and embryo quality. Embryos from both treated and control groups were transferred together in 20 transfer procedures, and in two couples fertilization was not obtained. Nine pregnancies occurred: one biochemical, one miscarriage, and seven patients delivered 9 healthy babies. Two pregnancies were twin with an overall pregnancy rate of 40.9% per cycle and of 45% per transfer. In conclusion, the results of the present study demonstrate that, in humans, extracellular ATP induces a significant increase of sperm fertilizing potential, as these findings are a rationale for the use of ATP for in-vitro treatment of human spermatozoa during IVF.

Adenosine Triphosphate↗
PubMed1986

Prematurely condensed human sperm chromosomes after in vitro fertilization (IVF).

During an in vitro fertilization (IVF) program 122 inseminated eggs showing polar body extrusion, but neither formation of pronuclei nor cell cleavage were analysed cytogenetically. Nine of these eggs showed prematurely condensed sperm chromosomes of the G1-phase (G1-PCC) besides the haploid set of maternal metaphase II chromosomes. This phenomenon can be explained by the permanent arrest of the oocytes at metaphase II after sperm penetration and hence the continuing presence of cytoplasmic chromosome condensing factors which lead to the induction of PCC in the sperm nucleus. The overall frequency of this aberrant type of fertilization was calculated to be in the order of 3-4% of all in vitro fertilized eggs.

Chromosome Banding↗
PubMed1986

Correlation between the zona-free hamster egg sperm penetration assay and human in vitro fertilization.

The association between in vitro fertilization (IVF) and the zona-free hamster egg sperm penetration assay (SPA) was studied in 134 couples. The indications for IVF were tubal disease in 82 couples, unexplained infertility in 23, male infertility in 10, and combined mechanical and male factors in 19. In general, a positive SPA was a good predictor of subsequent IVF (91 of 107 couples). Specificity (number of couples with fertilization in vitro divided by the number of couples with a positive SPA times 100) was 94% for tubal infertility, 76% for unexplained infertility, and 46% for male infertility. A negative SPA predicted an overall fertilization failure of 78% (21 of 27 couples). Sensitivity of the test (number of couples with IVF failure divided by the number of couples with a negative SPA times 100) was 100% for tubal and unexplained infertility but only 63% for male infertility. The high predictive value of the SPA for subsequent IVF outcome in tubal infertility and unexplained infertility warrants its routine use for prescreening in IVF programs. Because of the lack of association between SPA and IVF in oligoasthenospermia, the bioassay should not be relied upon for predicting IVF outcome in male subfertility. The overall high association between the animal model and human IVF reinforces the use of the SPA for both basic research and clinical decision-making.

Animals↗
PubMed1993

Effectiveness of in vitro maturation and in vitro fertilization techniques in pigs.

In vitro maturation and in vitro fertilization techniques in pigs have progressed considerably in recent years. Many reports focus on the factors affecting in vitro maturation that lead to normal male pronuclear formation or monospermy after fertilization in vitro. It is suggested that pig follicular fluid (pFF), follicle somatic cells and various hormones are important factors for the maintenance of cytoplasmic maturation of oocytes in vitro, but that fetal calf serum (FCS), which is generally added to maturation medium, is detrimental. A series of experiments clearly indicate that the glutathione (GSH) content of matured oocytes increases greatly when maturation medium is supplemented with cysteine, a precursor of GSH, and the rates of male pronuclear formation increase in parallel with the increasing GSH content. To prevent polyspermy, conditions of maturation and of fertilization in vitro are important. Culture of oocytes in medium with FCS for the first 24 h and with BSA for the second 24 h decreases the incidence of polyspermy, without a significant effect on nuclear maturation. However, it has been shown that secretory macromolecules of the oviduct may reduce the incidence of polyspermy by interacting with fertilizing spermatozoa rather than with oocytes. A reduction of polyspermy by treating spermatozoa with pFF is also reported. In addition to the many improvements in the methodology of in vitro fertilization using unfrozen spermatozoa in pigs, techniques for fertilizing oocytes in vitro with frozen epididymal and ejaculated spermatozoa have also recently been developed.

Animals↗
PubMed1991

The mandation of insurance coverage for in vitro fertilization.

For many infertile couples, in vitro fertilization represents a last hope; yet few couples can afford this expensive procedure, which health insurers do not routinely cover. To force health insurers to pay for in vitro fertilization, infertile couples have successfully lobbied in six state legislatures for "mandation" legislation, which will force insurers to pay for the procedure. This paper discusses the legislative battlefield for in vitro fertilization mandation and the long-term implications of such victories.

Bioethics↗
PubMed2006

[Use of biochemical markers to predict the outcome of pregnancies conceived by in vitro fertilization].

BACKGROUND: Pregnancies obtained after in vitro fertilization and embryo transfer are at increased risk for an adverse outcome compared with women who conceive naturally. Multiple gestations also occur more frequently after in vitro fertilization. Therefore, there is a need for markers that accurately detect the establishment of pregnancy and predict its outcome as early as possible, allowing for modification of monitoring and treatment if required. Ultrasound examination is part of the routine follow-up after in vitro fertilization, but a gestational sac is not reliably visible until 33-37 days after ovulation induction. As a result, there is an ongoing effort to find endocrine markers that can earlier detect the establishment of pregnancy and forecast its outcome. OBJECTIVE: The authors' aim was to assess the predictive value of the following potential serum markers, measured in the second week after embryo transfer in samples collected prospectively during the past ten years at the Division of Assisted Reproduction of their department: total beta-hCG (theoretical post-embryo transfer day 11 values, calculated from levels in two samples collected with a difference of two days, based on the mathematical model describing its exponential increase in early pregnancy), inhibin A, and CA-125. METHODS: Data of patients undergoing IVF or intracytoplasmic sperm injection and embryo transfer between 1995 and 2001 were analyzed. Establishment of pregnancy was assessed by measuring total beta-hCG concentrations in two serum samples collected between 8 and 16 days after ET with a difference of two days. Measurement of inhibin A and CA-125 levels was performed in the same samples. Logistic regression analyses were used to study the association of these serum markers and the number of retrieved oocytes and transferred embryos with pregnancy outcome. Receiver-operating characteristic (ROC) curves were constructed to identify optimal cutoff levels for outcomes and to assess overall predictive accuracy. RESULTS AND CONCLUSIONS: (1) Day 11 total beta-hCG can be used to compare hCG levels in samples from different sampling days and to predict early pregnancy losses and multiple ongoing pregnancies with high sensitivity and specificity. (2) Inhibin A concentrations are more accurate than day 11 hCG levels for predicting preclinical abortion after IVF but they have no advantage in forecasting ongoing or multiple ongoing pregnancies. (3) Prognostic accuracy of CA-125 measurements for the prediction of pregnancy as well as its outcome is inferior to that achieved with inhibin A.

Abortion, Spontaneous↗
PubMed1992

A simplified approach to in vitro fertilization.

A simplified protocol for in vitro fertilization treatment was developed using low-dose clomiphene for ovarian stimulation, timing of human chorionic gonadotropin administration based solely on sonographic criteria, in-office egg retrieval and use of a simplified culture medium for embryo culture. Twenty-six infertile women underwent 39 treatment cycles, with 33 egg retrievals resulting in 29 embryo transfers. Clinical pregnancy was obtained in six patients (20.7% per embryo transfer), for a rate comparable to that achieved with conventional in vitro fertilization treatment in an age-matched control group. This simplification of in vitro fertilization treatment protocols can lead to decreased patient stress and costs without a substantially lower success rate.

Adult↗
PubMed2005

Headaches in women undergoing in vitro fertilization and embryo-transfer treatment.

BACKGROUND: In vitro fertilization and embryo transfer has become a common therapeutic modality in modern fertility medicines. Treatment protocols are associated with exaggerated hormonal fluctuations in order to promote multiple follicular development and support of embryo survival. Estrogen is considered to be closely linked to migraine, and its extreme serum level fluctuations have been considered to trigger migrainous headaches. OBJECTIVE: To assess the prevalence, nature, and timing of headache attacks during the course of in vitro fertilization and embryo-transfer treatments. METHODS: We retrospectively evaluated the prevalence and the characteristics of headache among 98 women undergoing in vitro fertilization and embryo transfer therapy using a structured telephone interview. RESULTS: Out of the 98 evaluated patients 25 were migraine sufferers. Headache attacks during the in vitro fertilization and embryo transfer protocol were reported by 28.6% (n = 28) of the women and were significantly more prevalent among the migraineurs. The attacks were of debilitating severity in 82% (n = 23) of patients who reported headaches during treatment. Headache occurred most frequently following gonadotropin-releasing hormone (GnRH) analog induced hypothalamic-pituitary-ovarian axis downregulation, typically associated with very low 17-beta-estradiol (17-beta-E2) serum level. CONCLUSIONS: In vitro fertilization and embryo-transfer treatment might be associated with various degrees of headache. This is mostly observed in patients with migraine headache background and specifically during the downregulation stage of treatment, when very low levels of 17-beta-E2 are observed. (Headache 2005;45:215-219).

Embryo Transfer↗
PubMed2004

Natural-cycle in vitro fertilization combined with in vitro maturation of immature oocytes is a potential approach in infertility treatment.

OBJECTIVE: To describe pregnancies and live births that resulted from IVF of mature oocytes retrieved from dominant follicles in a natural cycle combined with in vitro maturation (IVM) of immature oocytes retrieved from small follicles. DESIGN: Case reports. SETTING: McGill Reproductive Center, Royal Victoria Hospital, McGill University. PATIENT(S): Three women with normal ovaries or polycystic ovaries who underwent infertility treatment. INTERVENTION(S): Administration of s.c. hCG (10,000 IU) 36 hours before oocyte retrieval in a natural cycle. After aspiration of all follicles, mature oocytes were inseminated immediately; immature oocytes were matured in vitro, inseminated by intracytoplasmic sperm injection (ICSI), and then the embryos transferred. MAIN OUTCOME MEASURE(S): Pregnancy and live birth. RESULT(S): Three pregnancies (two live births and one ongoing at time of writing) were achieved after the combination of natural-cycle IVF with IVM after transfer of the resulting embryos. CONCLUSION(S): Natural-cycle IVF combined with IVM might be a new approach to IVF treatment for women with various causes of infertility.

Adult↗
PubMed2006

Intracytoplasmic sperm injection (ICSI) in unexplained and stage I endometriosis-associated infertility after fertilization failure with in vitro fertilization (IVF).

PURPOSE: To investigate possible differences between unexplained and stage I endometriosis-associated infertility in ICSI cycles conducted after low fertilization (<20%) in preceding IVF cycles with normal semen parameters. METHODS: Retrospective cohort study consisting of patients with unexplained (n=48) and stage I endometriosis-associated infertility (n=43) with a minimum of one IVF cycle with <20% fertilized oocytes and normal semen quality, treated with ICSI from January 1997 to January 2006. Age matched male factor infertility patients (n=91) were used as controls. RESULTS: Diploid fertilization rate was significantly lower in the stage I endometriosis-associated infertility group compared to the unexplained infertility group. Score of the transferred embryos, implantation rate, pregnancy rate and outcome were similar in the two groups. CONCLUSIONS: ICSI appears to be an efficient treatment option after fertilization failure with IVF in unexplained and stage I endometriosis-associated infertility.

Adult↗
PubMed1989

Failed fertilization in human in vitro fertilization analyzed with the deoxyribonucleic acid-specific fluorochrome Hoechst 33342.

The degree and normality of nuclear maturation were assessed with the fluorochrome Hoechst 33342 in two groups of inseminated human oocytes that had failed to undergo fertilization. Group 1 consisted of 67 oocytes from 27 patients, each of whom had at least two other oocytes that had been fertilized and had cleaved. Group 2 consisted of 65 oocytes from 14 patients, none of whose oocytes had been fertilized. In group 1, 52.3% of the oocytes were found to be immature (germinal vesicle stage or metaphase-telophase I), whereas in group 2 only 26% were found to be immature. Thus oocyte nuclear immaturity was the major cause of fertilization failure when companion oocytes were fertilized. When no oocytes of a patient were fertilized, most oocytes were found to be mature, so other factors, such as sperm dysfunction or zona binding abnormalities, must account for most of the fertilization failure in this group of patients.

Benzimidazoles↗
PubMed2005

Intracytoplasmic sperm injection as a treatment for unexplained total fertilization failure or low fertilization after conventional in vitro fertilization.

OBJECTIVE: To determine whether IVF or intracytoplasmic sperm injection (ICSI) should be the choice of treatment in case of a previous IVF attempt with unexplained total fertilization failure or low fertilization (<25%). DESIGN: Prospective study. SETTING: Leiden University Medical Center. PATIENT(S): Thirty-eight couples undergoing IVF and ICSI on sibling oocytes after a first IVF attempt with total fertilization failure or with low fertilization (<25%). INTERVENTION(S): Performing IVF and ICSI on sibling oocytes. MAIN OUTCOME MEASURE(S): Fertilization and (ongoing) pregnancy rate. RESULT(S): A total of 271 oocytes were collected in 24 oocyte retrievals in the total fertilization failure group. Hundred nine oocytes were randomly allocated to IVF and 12 were fertilized (11%); 162 sibling oocytes were allocated to ICSI and 78 were fertilized (48%). In 8 of the 24 patients fertilization occurred after IVF. The pregnancy rate after transfer of 1 IVF and 1 ICSI embryo (n = 3) was 67% and after the transfer of 2 ICSI embryos (n = 21) this was 52%. In the low fertilization group 169 oocytes were collected in 14 oocyte retrievals. Seventy-two oocytes were randomly allocated to IVF and 16 were fertilized (22%). Ninety-seven sibling oocytes were allocated to ICSI and 58 were fertilized (60%). In 7 of 14 patients fertilization occurred after IVF. The pregnancy rate after the transfer of 1 IVF and 1 ICSI embryo (n = 5) was 80% and after the transfer of 2 ICSI embryos (n = 9) this was 33%. CONCLUSION(S): Performing ICSI on some oocytes of a cohort may avoid total fertilization failures both in patients with a history of total fertilization failure and in patients with a history of low fertilization, as the percentage of fertilization is higher after ICSI compared to IVF and the recurrence of total fertilization failure and low fertilization is high after IVF treatment.

Adult↗