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PubMed2004

Influence of sperm:oocyte ratio during in vitro fertilization of in vitro matured cumulus-intact pig oocytes on fertilization parameters and embryo development.

The present study was conducted to evaluate the influence of sperm:oocyte ratio during in vitro fertilization (IVF) of in vitro matured cumulus-intact oocytes on fertilization parameters and embryo development in pigs. In vitro matured oocytes surrounded by intact cumulus cells (COC) were inseminated with frozen-thawed spermatozoa at different sperm:oocyte ratios (2000:1, 3000:1, 4000:1, 6000:1, and 8000:1). Denuded oocytes inseminated with 2000 frozen-thawed spermatozoa:oocyte were the control group. A total of 2546 oocytes in five replicates were exposed to spermatozoa for 6 h and then cultured in embryo culture (EC) medium for 6 h (pronuclear formation) or 7 days (blastocyst formation: BF). The penetration rate increased in the COC groups with the sperm:oocyte ratio, reaching the highest rates with 8000:1 spermatozoa:oocyte (72.1 +/- 6.5%), similar to the control (73.5 +/- 3.5%). However, the monospermy was highest with the lower spermatozoa:oocyte rates (82.6-94.8%) and decreased drastically (P<0.05) in the COC group fertilized with 8000 sperm:oocyte (36%). The efficiency of fertilization (number of monospermic oocytes/total number of inseminated oocytes) showed no difference among the COC groups (20-30%) but they were significantly lower (P<0.007) than those obtained by the control group (43.7 +/- 2%). Embryo development was highest in the control group (58% for cleavage and 23% for BF) but not significantly different with the 6000 and 8000 sperm:oocyte COC groups (47 and 50% for cleavage and 19 and 17% for BF, respectively). These results indicate that the use of COC for IVF involves a drop in the efficiency of the fertilization and the necessity to increase the frozen-thawed sperm:oocyte ratio three to four times more to obtain similar embryo development to denuded oocytes.

Animals↗

PubMed1975

In vitro fertilization of rat and mouse eggs by ejaculated sperm and the effect of energy sources on in vitro fertilization of rat eggs.

In vitro fertilization of rat and mouse eggs by ejaculated or epididymal spermatozoa in chemically defined media was studied. Penetration rates by ejaculated sperm was very low (0 to 8%) in the rat, but 11 to 41% of eggs were penetrated by ejaculated sperm in the mouse. The optimal concentration of sperm for in vitro fertilization appears to be similar whether ejaculated or epididymal sperm were used. The time of sperm penetration in the mouse eggs, however, was delayed for one-half to one hour when ejaculated sperm were used. The importance of sodium pyruvate, sodium lactate and glucose in the medium containing bovine serum albumin for in vitro fertilization of rat eggs was examined. When rat eggs in cumulus clot were exposed to epididymal sperm preincubated for five hours, the presence of sodium pyruvate, sodium lactate and glucose was found to play an important role. When exposed to non-incubated epididymal sperm sodium pyruvate could be omitted without much decline of the fertilization rate. When the denuded eggs were exposed to non-incubated sperm, penetration rates were very low (0 and 5%) in the absence of pyruvate. It appears that although lactate, pyruvate and glucose are all important for in vitro fertilization of rat eggs, pyruvate can be supplied by the follicular cells surrounding the eggs.

Animals↗

PubMed2002

A hierarchical Bayesian approach to modeling embryo implantation following in vitro fertilization.

In vitro fertilization and embryo transfer (IVF-ET) is considered a method of last resort for treating infertility. Oocytes taken from a woman are fertilized in vitro, and one or more resulting embryos are transferred into the uterus, with the hope that at least one will implant and result in pregnancy. Successful implantation depends on both embryo viability and uterine receptivity. This has led to the development of the EU model for embryo implantation, wherein uterine receptivity is characterized by a latent binary variable U and embryo viability is characterized by a latent binomial variable E representing the number of viable embryos among those selected for transfer. The observed number of implantations is the product of E and U. Zhou and Weinberg (1998) developed a regression formulation of the EU model in which embryo viabilities are independent within patients. We extend their methodology to a Bayesian hierarchical framework that allows for correlation between the embryo viabilities and gives explicit characterization of patient-level heterogeneity. When some subjects have zero implantations, the likelihood for the hierarchical EU model is relatively flat and therefore using prior information for key parametersis needed. This provides a key motivation for adopting a Bayesian approach. The model is used to assess the effect of hydrosalpinx on embryo implantation in a cohort of 288 women undergoing IVF-ET because of tubal disease. Hydrosalpinx is a build-up of fluid in the Fallopian tubes, which sometimes leaks to the uterus and may reduce the likelihood of implantation. The EU model is well suited to this question because hydrosalpinx is thought to affect implantation by reducing uterine receptivity only. Our analysis indicates substantial subject-level heterogeneity with respect to embryo viability, suggesting the utility of a multi-level model.

Journal Article↗

PubMed1993

Intermediate filament protein expression in early developmental stages of the mouse. A confocal scanning laser microscopy study of in vitro fertilized and in vitro cultured pre-implantation mouse embryos.

Expression patterns of intermediate filament proteins have been studied during early mouse embryo development. For this purpose, pre-implantation embryos at different stages of development after in vitro fertilization were studied using antibodies to cytokeratins, vimentin and lamins, using the indirect immunofluorescence assay. The levels of expression were quantitated and localization of the protein constituents was assessed by means of confocal scanning laser microscopy. Our studies showed that, although the embryos grew in culture, vimentin could not be detected in a filamentous organization. Immunofluorescence for cytokeratins was only positive from the 8-cell stage onwards. In the morula stage an increased level of cytokeratin expression was observed with a transitional staining pattern, combining a filamentous and a diffuse occurrence. In the blastocyst stages profound cytokeratin filaments were seen in trophoblast cells but not in the inner cell mass. When the cytokeratin subtypes were analysed separately, it became apparent that expression levels of cytokeratins 8 and 18 increased gradually up to a filamentous pattern in the blastocyst stage. Cytokeratins 7 and 19, although elevated in the latter stage and showing a filamentous distribution, were not found as prominently as cytokeratins 8 and 18. A-type as well as B-type lamins could be detected in all developmental stages examined, as a faintly reactive nuclear lamina. In blastocysts both lamin types were detected in trophoblast as well as in inner cell mass.

Animals↗

PubMed1985

Follicular development: lessons learned from human in vitro fertilization.

In vitro fertilization has offered new insights into our understanding of ovulation induction, folliculogenesis, and luteal phase events. This new information is provided by the ability to precisely study these cycles in a frequent and sequential fashion through the use of peripheral blood markers, ultrasound evaluation, and follicular fluid constituents and cell culture techniques, as well as direct observation of the oocyte, fertilization, and cleavage. In these stimulated cycles the follicular phase serum estradiol levels in conjunction with ultrasound were evaluated; a poor correlation was shown between follicle size and number and estrogen production. This distinct dyssynchrony suggests the recruitment of a number of cohorts of follicles in each stimulated cycle. From the biochemical markers in follicular fluid, cyclic adenosine monophosphate has a distinct predictive value in regard to pregnancy in in vitro fertilization-embryo transfer cycles. In the luteal phase, the mass effect of aspiration of great numbers of granulosa cells, the effect of supplemental progesterone, and the influence of high follicular phase estradiol levels remain controversial and, therefore, a less clear cut pattern emerges. Variations in the protocol have not greatly improved the major problems of folliculogenesis associated with ovulation induction and an in vitro fertilization-embryo transfer program, that is, follicular asynchrony and luteal phase deficiency.

Body Fluids↗

PubMed1996

Effect of hypotaurine on in vitro fertilization and production of term offspring from in vitro-fertilized ova of the Japanese field vole, Microtus montebelli.

The present study was proposed to establish new in vitro fertilization (IVF)-embryo transfer techniques for wild mammals. We demonstrated that IVF of the Japanese field vole, Microtus montebelli, is improved by hypotaurine, and that IVF vole embryos can develop to normal offspring after transfer to recipient females. The rate of IVF was significantly increased when 0.1 or 1 mM of hypotaurine was added during sperm preincubation and fertilization (p < 0.05). Preincubation of spermatozoa with 1 mM hypotaurine prior to oocyte insemination did not have a significant effect on fertilization rate (p < 0.01). At 96 h after insemination, most cultured embryos developed to the 2-cell stage (68-83%), but development to blastocysts was very low (0-3%) for all treatments. Although 43 living offspring were produced after transfer of IVF pronuclear embryos to recipients, the delivery rate of recipients was very low (21%). The litter size of pseudopregnant recipients that produced a litter was 4-11, and the rate of in vivo development of transferred embryos in the pregnant recipients was 41%. These results suggest that hypotaurine affects IVF rather than sperm preincubation and that IVF embryos can develop to normal offspring.

Animals↗

PubMedDate not supplied

[Results after an in vitro fertilization pregnancy. French National Register on In Vitro Fertilization].

For this analysis, all the oocyte recoveries realised between 1989, January 1st and 1995, December 31th and collected by the French national register on IVF (FIVNAT) were included. Three groups were constituted, depending on the existence of a previous birth (group 1, n = 8,615), or of a previous failed pregnancy (spontaneous abortion or ectopic pregnancy, group 2, n = 8,539), both obtained by IVF, or no previous IVF pregnancy (group 3, n = 139,843). In this last group, women and men were of younger age (respectively 33.3 +/- 4.7 vs 35.1 +/- 3.9 for group 1 and 34.3 +/- 4.5 for group 2, p < 0.001, and 35.2 +/- 5.9 vs 37.0 +/- 5.1 for group 1 and 36.1 +/- 5.6 for group 2, p < 0.001). Overall results were lower in the group 3. The average number of embryos was 3.7 +/- 3.7 compared to 4.4 +/- 3.8, p < 0.001 for the two other groups. The fertilization rates was at 47.3 +/- 33.2% in group 3, compared to 55.5 +/- 30.5 in group 1 and 55.6 +/- 30.4 in group 2 (p < 0.001). Finally, the per recovery pregnancy rates were respectively 17.9%, 28.0%, 25.0% (p < 0.001). In group 3, infertility origin was less often due to a tubal disease (53.2% vs 65.6% in group 1 and 66.6% in group 2, p < 0.001). On the opposite, an isolated male origin was more frequent (respectively 19.9%, 11.0% and 12.2%) (p < 0.001). In group 3, compared with those with secondary infertility, women with primary infertility were younger (32.6 +/- 4.6 vs 34.7 +/- 4.6, p < 0.001), had more oocytes (8.9 +/- 5.7 vs 8.1 +/- 5.4, p < 0.001), but less embryos (3.6 +/- 3.7 vs 3.9 +/- 3.7, p < 0.001), a decreased fertilization rate (44.8 +/- 33.4 vs 51.6 +/- 32.6, p < 0.001) and a decreased pregnancy rate (17.3% vs 18.9%, p < 0.01). Infertility was more often of male origin (23.6% vs 11.7%, p < 0.001).

Adult↗

PubMed1986

The treatment of infertility associated with endometriosis by in vitro fertilization.

In vitro fertilization and embryo transfer (IVF-ET) was performed on women with tubal disease only (28 women, 40 cycles), endometriosis grade I (15 women, 24 cycles), grade II (25 women, 37 cycles), grade III (26 women, 36 cycles), or grade IV (31 women, 52 cycles). Rates of oocyte recovery and fertilization were not affected by the presence of endometriosis. Pregnancy rates per cycle were similar to that of the tubal group (18%) for women with grade I (13%) or grade II (14%) endometriosis, lower in women with grade III endometriosis (6%) and significantly reduced in women with grade IV (2%, P less than 0.05).

Clomiphene↗

PubMed1987

Superovulation with intrauterine insemination in the treatment of infertility: a possible alternative to gamete intrafallopian transfer and in vitro fertilization.

In vitro fertilization and embryo transfer (IVF-ET) and gamete intrafallopian transfer (GIFT) are used to treat intractable infertility in women with no distortion of the pelvic viscera, despite the lack of controlled trials demonstrating efficacy. The mechanism of any purportedly enhanced cycle fecundity in ovulatory women without significant distortion of the pelvic viscera is unclear, but both GIFT and IVF-ET increase the number of male and female gametes at the site of fertilization. Intrauterine insemination (IUI) during human menopausal gonadotropin (hMG)-stimulated superovulatory cycles has similar potential but does not require oocyte retrieval. To evaluate the possibility that simply increasing the number of gametes at the site of fertilization might account for pregnancies attributed to IVF-ET and GIFT, the authors retrospectively analyzed the outcome of couples undergoing IUI during hMG cycles between 1983 and 1986 in women with normal pelvic anatomy. IUI during hMG-stimulated cycles yielded a cycle fecundity (f) of 0.17 for endometriosis, 0.29 for cervical factor, and 0.19 for idiopathic infertility, which approaches the fecundity of normal women and equals or exceeds that reported for IVF-ET and GIFT. The authors conclude that treatment with IUI in hMG cycles, alleviating the need for invasive oocyte retrieval, should be considered for inclusion in a randomized, controlled trial in comparison with IVF-ET and GIFT.

Cervix Mucus↗

PubMed1989

Improved pregnancy outcome with the addition of leuprolide acetate to gonadotropins for in vitro fertilization.

In vitro fertilization treatment outcomes were compared prospectively in unselected patients with and without the addition of leuprolide acetate to gonadotropins for ovarian hyperstimulation. While the leuprolide patients required greater quantities of exogenous gonadotropins to achieve ovarian stimulation, significant improvements in the fertilization, implantation, and spontaneous abortion rates were observed compared with patients not receiving leuprolide. The ongoing pregnancy ("take home baby") rates per aspiration with and without leuprolide were 29% and 12%, respectively. Furthermore, the multiple pregnancy rate was markedly increased in the leuprolide group (44% versus 8%), suggesting that the improved pregnancy outcome with pituitary suppression was due primarily to higher oocyte and embryo quality.

Adult↗

PubMed1999

Social and ethical aspects of in vitro fertilization.

In vitro fertilization (IVF) stands out as one of the contemporary period's most extraordinary technologies, and its social and ethical consequences among the most far reaching. Despite its uncertain effectiveness and medical consequences, IVF has contributed significantly to the medicalization of infertility and the increasingly imperative character of reproductive technology. New developments in IVF, particularly oocyte donation, have created new definitions of treatable infertility and new social needs for IVF; when the technology does not result in pregnancy or healthy babies, these developments have created profound new disappointments. IVF and the commodification of the extracorporeal embryo have also confused the social meaning and legal definition of parenthood. Ultimately the relationship between prospective parents, infertility specialists, and the embryos that they create is a highly ambiguous one. This ambiguity is likely to be a long-term characteristic of efforts to develop, use, and assess assisted reproductive technologies.

Commodification↗

PubMed1997

Assisted reproduction and reproductive rights: the case of in vitro fertilization.

In vitro fertilization (IVF) and other assisted reproduction technologies (ARTs) have become widely accepted as therapy for a wide array of fertility problems and accompanied by the rapid expansion of clinics that provide full range of ARTs. Although these technologies undoubtedly offer benefits for some individuals, they raise important questions over reproductive rights to safe and effective treatment as well as access. This article analyzes current data concerning the safety, effectiveness, and cost of IVF. It concludes that IVF and related techniques have been transformed too rapidly and easily from experimental to therapy status, despite evidence that suggests considerable caution is warranted. Unfortunately, the widespread diffusion of IVF has preceded rather than followed firm evidence of its value in extending the reproductive rights of women and couples. Resources might better be directed toward prevention of fertility problems and discovering the causes of infertility.

Cost-Benefit Analysis↗

PubMed2001

Cost-effectiveness of in vitro fertilization.

In vitro fertilization (IVF) is an effective but underutilized treatment for long-standing infertility. The median projected cost per IVF cycle in 2001 in the United States would be $9226 and $3531 in 25 other countries, based on previously published estimates. The cost per delivery arising from IVF cycles in 2001 in the United States would average $56,419, and $20,522 in eight other countries, based on previously published estimates. The cost-effectiveness of IVF has not been proven in three randomized controlled trials, but these trials are difficult to design and implement. Multiple gestation births significantly increase the cost of IVF treatment. IVF multiple births cost 36% more than the IVF cycles that gave rise to the multiple births. The uptake of IVF in most countries falls short of the minimum 1500 cycles per annum that would be needed per million population. Comparative international data indicate that infertile couples would increase their uptake of IVF services if the price were lower.

Adult↗

PubMedDate not supplied

Couples' experiences with in vitro fertilization.

In vitro fertilization (IVF) is one of the newest techniques for treatment of infertility. While the medical aspects of IVF have been extensively reported, little research has explored this procedure from the couple's point of view. In this descriptive study, 28 couples who had experienced at least one IVF procedure were interviewed. The results indicate that while IVF offers hope for infertile couples, the procedure, if unsuccessful, can be emotionally traumatic. In addition, the results suggest that comprehensive anticipatory information and emotional support are the primary needs of couples undergoing IVF.

Adaptation, Psychological↗

PubMed1983

[Collection of mature ovocytes during cycles with spontaneous ovulation for the purpose of in vitro fertilization].

In vitro fertilization was attempted during a spontaneous cycle in 17 out of 97 patients. Preovulatory urinary estrogen and LH maximas were assayed in order to time celioscopy 24 hours after the onset of the triggering urinary LH maxima. In 8 patients, comparison with plasmatic LH shows that the timing of spontaneous ovulation can be precisely predicted from urine assays. However, when the triggering LH release is weak, LH must be above or equal to 25 IU/g of creatinine with a creatininuria above or equal to 0.4 g/l in two urine samples at a three-hour interval.

Estrogens↗

PubMed1995

Cost analysis of in vitro fertilization.

In vitro fertilization (IVF) has become a routine tool in the arsenal of infertility treatments. Assisted reproductive techniques are expensive, as reflected by the current "take home baby" rate of about 15% per cycle, implying the need for repeated attempts until success is achieved. Israel, today is facing a major change in its health care system, including the necessity to define a national package of health care benefits. The issue of infertility and whether its treatment should be part of the "health basket" is in dispute. Therefore an exact cost analysis of IVF is important. Since the cost of an IVF cycle varies dramatically between countries, we sought an exact breakdown of the different components of the costs involved in an IVF cycle and in achieving an IVF child in Israel. The key question is not how much we spend on IVF cycles but what is the cost of a successful outcome, i.e., a healthy child. This study intends to answer this question, and to give the policy makers, at various levels of the health care system, a crucial tool for their decision-making process. The cost analysis includes direct and indirect costs. The direct costs are divided into fixed costs (labor, equipment, maintenance, depreciation, and overhead) and variable costs (laboratory tests, chemicals, disposable supplies, medications, and loss of working days by the couples). The indirect costs are the costs of premature IVF babies, hospitalization of the IVF pregnant women in a high risk unit, and the cost of complications of the procedure. According to our economic analysis, an IVF cycle in Israel costs $2,560, of which fixed costs are about 50%. The cost of a "take home baby" is $19,267, including direct and indirect costs.

Costs and Cost Analysis↗

PubMed1996

Application of image analysis cytometry in follicular fluid cells obtained from in-vitro fertilization cycles: relationships to patient's age, oocyte maturity, fertilizability and in-vitro fertilization outcome.

In an in-vitro fertilization (IVF)/embryo transfer programme granulosa cells obtained from 59 individual preovulatory follicles were analysed using multiparameter image analysis cytometry, in an attempt to determine whether their morphometric and DNA-cytometric parameters could prove useful in assessing follicle and oocyte maturity and in predicting fertilizability and outcome of these IVF cycles. Almost all morphometric and DNA-cytometric parameters were not correlated with either the patient's age or oocyte maturity, and did not predict oocyte fertilization or occurrence of a clinical pregnancy. The only possible relevant parameter which, despite its inverse correlation to total luteinizing hormone administration, also proved to be inversely correlated to pregnancy outcome (in the seven cases in which a pregnancy occurred), was the percentage of granulosa cell nuclei with increased DNA content (> 5c). Finally, if granulosa cells do not reveal euploid polyploidization in spontaneous or induced ovulatory cycles, the detected cells with increased DNA content should be interpreted as aneuploid, i.e. with chromosomal aberrations, and so their presence could also be discussed in connection with the hypothetical risk of prospective neoplastic transformation of the tissue.

Adult↗

PubMed2005

Sperm binding, in vitro fertilization, and in vitro embryonic development of bovine oocytes fertilized with spermatozoa incubated with norepinephrine.

The final stages of sperm maturation, fertilization, and early embryonic development occur within the oviduct and are essential for successful reproduction in mammals. Norepinephrine was previously identified in native bovine oviductal fluid and its in vitro effects on bull sperm capacitation and the acrosome reaction have been determined. It was unknown how physiological concentrations of norepinephrine influence sperm binding, fertilization, and embryo development. Therefore, the objective of this study was to determine if pre-incubating bovine spermatozoa with physiological concentrations of norepinephrine prior to insemination of bovine oocytes would improve sperm-oocyte binding, fertilization, and embryonic development in vitro. Norepinephrine, in concentrations representing those measured in bovine oviductal fluid, was used to treat bovine spermatozoa prior to insemination. Spermatozoa incubated in norepinephrine were used to inseminate bovine oocytes matured in vitro, and oocytes were evaluated for sperm binding and fertilization. Additional experiments were conducted to evaluate how early in the co-incubation period oocytes were fertilized by spermatozoa pre-incubated with norepinephrine, and to test the developmental competence of those oocytes fertilized with norepinephrine-treated sperm. Sperm binding to the zona pellucida was reduced by pre-incubation with norepinephrine. Rates of fertilization and embryo development did not increase as a result of pre-incubating spermatozoa with norepinephrine, but as early as 4h after insemination, spermatozoa treated with 20 ng/ml norepinephrine fertilized more oocytes than spermatozoa incubated in medium alone. Interestingly, this concentration of norepinephrine was found to capacitate spermatozoa in previous studies. These data suggest that oocytes fertilized by spermatozoa incubated in 20 ng/ml norepinephrine fertilize earlier in vitro than sperm pre-incubated in medium alone, and provide additional support for the role of norepinephrine in sperm capacitation and the acrosome reaction.

Animals↗