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Results for “In Vitro Fertilization (IVF)”

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PubMed2003

The proportion of follicular fluid CD16+CD56DIM NK cells is increased in IVF patients with idiopathic infertility.

One-fifth of all in-vitro fertilization (IVF) patients suffer from idiopathic infertility. A low fertilization rate is one of the most characteristic features of IVF in this group, probably caused by oocyte dysfunction. We speculate that an altered lymphocyte profile in follicular fluid (FF) may affect oocyte function and thus play a role in idiopathic infertility. Therefore, we compared levels of lymphocyte populations present in FF of 11 patients with idiopathic infertility (study group) with 29 patients in the control group, i.e. severe male factor infertility (n=17) or tubal factor infertility (n=12). Triple color flow cytometry was used to discriminate between T cells and NK cell subpopulations. In the idiopathic infertility group, a shift from T to NK cells was observed in FF as compared to the control group, caused mainly by a significant higher level of NK cells--20.3 and 13.6% (P<0.05), respectively. This high level of NK cells was due to a rise of the CD16+CD56dim NK cell subset. In peripheral blood, the NK cell levels showed a similar although not significant trend (P=0.08). As the CD16+CD56dim NK cell subpopulation is known for its cytotoxic properties, this subpopulation may negatively affect folliculogenesis and oocyte maturation, reflected by a diminished fertilization rate in the idiopathic infertility group. An altered lymphocyte profile in FF could therefore influence fertility in these patients.

B-Lymphocytes↗

PubMed1987

Uterine evaluation by microhysteroscopy in IVF candidates.

Over an eight-month period, microhysteroscopy was performed on 180 women, candidates for in-vitro fertilization (IVF). One hundred and two of them were suspected of having uterocervical pathology (group I), and 78 were not (group II). Group I had microhysteroscopic abnormalities in 60.8% and group II in 21.8%. Microhysteroscopy showed 30.6% of 36 positive hysterographies to be false positive and 37.5% of 144 negative hysterographies to be false negative, producing a confirmation rate of 63.9%. Admission to the IVF programme depended on microhysteroscopic normality. One hundred and one women were accepted directly, and 20 refused ('primary decision'). Fifty-nine were deferred with treatable hysteroscopic abnormalities, pending medical (22) and surgical (37) treatment, 14 via the microhysteroscope. Of the 59 women deferred, 51 were finally admitted and 8 rejected after a microhysteroscopic check ('secondary decision'). Finally, 152 of the 180 candidates (84.4%) were admitted. Microhysteroscopy helped to determine the optimal route for embryo transfer in 23 women with cervical pathology. Term pregnancy rates per embryo transfer were similar with hysteroscopic findings primarily normal, or normal after successful treatment (11.2 and 10.8% respectively). Microhysteroscopy seems useful for selection of cases for IVF, accurately determining utero-cervical pathology or normality, aiding in surgical correction of abnormalities and facilitating the process of embryo transfer. We suggest that microhysteroscopy should be performed routinely on all IVF candidates.

Adult↗

PubMed1996

Cytogenetics of uncleaved oocytes and arrested zygotes in IVF programs.

PURPOSE: Cytogenetic studies of arrested oocytes and zygotes were used to understand in vitro fertilization (IVF) failures. METHODS: We investigated the cytogenetics (Giemsa banding and FISH) of 710 uncleaved oocytes and 94 arrested zygotes from 208 patients undergoing IVF procedures. RESULTS AND CONCLUSIONS: Of uncleaved oocytes without a polar body, 39% were judged cytogenetically abnormal (17% unbalanced predivision and 21.5% diploid). Of 575 oocytes with a polar body, 124 (21.5%) showed numerical or structural chromosome aberrations. In arrested zygotes, approximately equal cases were found with separate condensed haploid complements (no syngamy), nuclear asynchrony and pulverized DNA, and apparently cytogenetically normal zygotes arrested at mitosis. These data on chromosome abnormalities were also analyzed with respect to two ovarian stimulation protocols and to maternal age. Both ovarian stimulation protocols and to maternal age. Both ovarian stimulation protocols showed the same levels of chromosome abnormalities. Overall chromosome abnormalities and premature chromosome condensation were also unchanged with maternal age. These data illustrate the significance of chromosome aberrations in IVF failures.

Adult↗

PubMed1989

The human blastocyst produces a soluble factor(s) that interferes with lymphocyte proliferation.

Animal studies have reported that the blastocyst produces immunosuppressive factors that are critical to successful implantation. The production of such factor(s) by the human blastocyst has not yet been described. To test that hypothesis, the spent media of 67 human embryos developed in vitro was evaluated using either the allogeneic one-way mixed lymphocyte response (MLR) or the concanavalin A (Con-A) stimulated lymphocyte response (SLR). Conditioned embryo media was obtained at in vitro fertilization (IVF) in aliquots of (A) the first 24 hours in vitro, (B) the second 24 hours in vitro, and (C) from 24 hours until development of the blastocyst stage. Control media consisted of Ham's F-10 medium (Gibco, Grand Island, NY) with identical maternal serum concentration to test media. Conditioned media (either A, B, or C) or control media was added to 200-microL wells containing lymphocytes in either the MLR or SLR. A significant suppressive effect of 25% to 60% was observed in seven of nine samples of blastocyst-conditioned media (P less than 0.05). No significant suppressive effect was found in the earlier embryo media (A or B) in either the MLR or the SLR. These data suggest that the human blastocyst produces a factor(s) that interferes with lymphocyte proliferation.

Blastocyst↗

PubMed2002

Diagnostic dilemma encountered when detecting bovine viral diarrhea virus in IVF embryo production.

Routine quality controls in production of bovine embryos by in vitro fertilization (IVF) should include screening all materials of animal origin for the presence of bovine viral diarrhea virus (BVDV). Using a reverse transcription nested polymerase chain reaction (RT-nPCR) assay, we detected BVDV in primary cultures of uterine tubal cells (UTC) that had been used during IVF procedures. The goal of our ensuing investigation was to determine its source and assess risks associated with the identified contaminant. Sequencing of the amplified 5' nontranslated region (NTR) of the viral genome confirmed a Genotype I BVDV contaminant. This viral contaminant was also identified by RT-nPCR in multiple samples of the same lot of fetal bovine serum (FBS) that was used in transport media by the laboratory that harvested the UTC. Both routine and enhanced roller bottle methods for virus isolation failed to detect BVDV in the FBS. Furthermore, virus neutralization assays did identify antibodies to Genotype I strains of BVDV in the FBS. After 7 days of co-incubation, neither cultured, washed UTC nor exposed, washed embryos were RT-nPCR positive for BVDV. Eight embryos produced in the contaminated system were nonsurgically transferred into eight seronegative cows. None of the embryo recipients seroconverted to BVDV. Thus, contamination of cell culture medium with BVDV did not result in transmission of the virus when IVF embryos were transferred. Failure to transmit disease was likely aided by serendipitous control from anti-BVDV antibodies in the FBS. However, a diagnostic dilemma was created when the RT-nPCR assays used to screen for BVDV were positive, yet attempts to isolate the virus were negative. This case study illustrates that if molecular assays are to be used to confirm the pathogen-free status of IVF embryo production systems, media components of animal origin (e.g. FBS) should be screened with molecular assays for BVDV as well as traditional virus isolation techniques.

5' Untranslated Regions↗

PubMed1992

Changes in the properties and composition of zona pellucida of pigs during fertilization in vitro.

Several hundred fertilized pig eggs were prepared by an in vitro fertilization (IVF) technique in which follicular phase ovarian eggs were matured in vitro to metaphase II before incubation with capacitated epididymal spermatozoa for 12 h at 39 degrees C. Parthenogenetic eggs were also prepared by stimulation of the mature eggs with an electric pulse. The zonae were solubilized with 0.2% pronase/phosphate-buffered saline (PBS) or lactic acid/PBS. The time taken for solubilization was 30-40% shorter than for unfertilized eggs, indicating that zona hardening was induced during fertilization. At the same time, the sperm receptor activity of the zona was reduced. Electrophoretic analyses of zona glycoproteins from the ovarian, mature and fertilized eggs revealed that the amount of 90 kDa proteins decreased substantially during fertilization. This fraction could barely be detected in the zonae from parthenogenetic eggs. However, modification with a fluorescent probe showed that the general architecture of the zona remained unchanged during fertilization. These results suggest that the minor 90 kDa proteins are specifically degraded by the protease(s) released from the oocyte at fertilization, thereby leading to the block to polyspermy.

Animals↗

PubMed2003

Effect of the volume of medium and number of oocytes during in vitro fertilization on embryo development in pigs.

The present study was designed to determine the effect of the volume of medium (VM) and the number of oocytes (NOOC) during in vitro fertilization (IVF) on embryo development in pigs. Groups of 15, 30 and 50 in vitro matured oocytes were transferred to 2, 1 and 0.1 ml of modified Tris-buffered medium (mTBM) and inseminated with frozen-thawed spermatozoa (2000 spermatozoa/oocyte) in a 3 x 3 factorial experiment. A total of 2739 oocytes from four replicates were exposed to spermatozoa for 6 h and then cultured in embryo culture medium for 6 h (pronuclear formation) or 7 days (blastocyst formation: BF). The efficiency of fertilization (EF: number of monospermic oocytes/total number of inseminated oocytes) and BF decreased (P<0.03) as the VM increased (EF: 45.9+/-2.2, 43.8+/-2.6 and 36.9+/-1.6% and BF: 29.4+/-2.7, 23.2+/-1.8 and 19.9+/-2.1% for VM 0.1, 1 and 2 ml, respectively). The BF, but not EF, was also affected (P<0.04) by NOOC (19.8+/-1.6, 28.1+/-2.3 and 24.6+/-2.9% for groups of 15, 30 and 50 oocytes, respectively). The effect of the interaction VM x NOOC on EF and BF was not significant. These results indicate that when 2000 spermatozoa/oocyte were used, a low volume of IVF medium (0.1 ml) and the number of oocytes during IVF (30-50) can improve the in vitro embryo production in pigs.

Animals↗

PubMed1998

Effect of maternal age and conditions of fertilization on programmed cell death during murine preimplantation embryo development.

One of the major morphological anomalies observed in many human pre-embryos is extensive cellular fragmentation. Previously we confirmed that embryo fragmentation seemed to be associated with the activation of programmed cell death (PCD). The purpose of our experiments was to establish a rate for murine embryo fragmentation in vivo after hormonal stimulation in young versus older females and to compare it with the rate of embryo fragmentation during in-vitro fertilization (IVF). While murine maternal age beyond 40 weeks increased the rate of embryo fragmentation following in-vivo fertilization (P = 0.001), oocytes from females of all ages had a uniformly high rate of fragmentation when fertilized in vitro (33%). None of the fragmented murine embryos proceeded further in development. In the mouse, fragmentation occurs exclusively during the first cell cycle. Furthermore, IVF significantly reduced the rate of blastocyst formation (P = 0.0001) and decreased the mean cell number at the blastocyst stage in comparison with embryos produced in vivo (P < 0.0001). The cell death index was significantly affected by both maternal age (P = 0.005) and IVF (P = 0.0001). Identification of specific factors which trigger PCD, especially those associated with IVF, may enable us to lower the rates of fragmentation in preimplantation embryos and thereby increase pregnancy rates after human IVF.

Animals↗

PubMed1993

Lesbian motherhood and genetic choices.

Rhonda, a physician, and Clara, a registered nurse, have been lovers for the past 7 years. Both women are in their 30s, in good health, and financially stable. Clara has contacted Dr. Robinson, a highly regarded obstetrician and geneticist, with an unusual request. Clara is seeking to become pregnant by means of in vitro fertilization (IVF) with an embryo transfer (ET) technology. She proposes being implanted with ova harvested from her partner, Rhonda, and fertilized in vitro with donor sperm from a local medical-school sperm bank. Dr. Robinson explains the relatively low success rate involved in IVF and ET techniques and suggests artificial insemination of Clara's own ova. Clara and Rhonda understand the low success rates but are undeterred. Their mutual goal is not merely to have a child but to share in the maternity of their children. What ethical issues are raised by the plan Clara and Rhonda propose? What are Dr. Robinson's ethical responsibilities with respect to the request by Clara and Rhonda?

Child↗

PubMed1992

Evaluating an in vitro culture system of bovine uterine and oviduct epithelial cells for subsequent embryo co-culture.

Three experiments were conducted to evaluate the effects of culture medium and incubation temperature on bovine uterine and oviduct epithelial cell growth, so that the most efficient combination could then be used to develop a co-culture system for bovine embryos. In the first experiment, uterine and oviduct epithelial cells at either the second or third subpassage were incubated for 8 days at 37 degrees C with 5% CO2 in Tissue Culture Medium-199, CMRL-1066, Minimal Essential Medium, Ménézo's B2 or Ham's F-12 medium. In addition to plotting growth curves of cell populations, the cell cycle was monitored for 8 days by flow cytometry. Uterine and oviduct epithelial cells incubated in CMRL-1066 exhibited the highest growth rates during the 8-day culture period. However, there were no differences in cell cycle analysis among treatment groups during the incubation period. In the second experiment, CMRL-1066 medium was used to evaluate growth and proliferation of uterine and oviduct epithelial cells incubated at 37 degrees C or 39 degrees C; temperature had no significant effect on growth rates or proliferation rates for either uterine or oviduct cells during the 8-day incubation. In the third experiment, the more promising culture media for epithelial cell culture studies were chosen for in vitro maturation and subsequent in vitro fertilization (IVF) of bovine oocytes. Early cleavage-stage embryos produced by IVF procedures were subsequently cultured in vitro for 7 days in medium alone or with oviduct epithelial cells. In this study, the culture medium did not influence fertilization or cleavage rates. However, more embryos co-cultured with oviduct epithelial cells were considered viable after 7 days of incubation compared with embryos incubated in medium alone. These results indicate that various incubation conditions can influence the growth of bovine uterine and oviduct epithelial cells in vitro. However, in spite of changes in cell growth patterns, there does not appear to be a change in their embryotropic capabilities in vitro.

Animals↗

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↗

PubMed1997

Outcome of IVF in DES-exposed daughters: experience in the 90s.

PURPOSE: The outcome of in vitro fertilization (IVF) in a group of infertile women with a history of in utero exposure to diethylstilbestrol (DES) was analyzed. Records from an academic IVF program were retrospectively reviewed. METHODS: Seventeen infertile women with a self-reported history of exposure to DES in utero, attending the IVF unit at Massachusetts General Hospital (MGH) for assisted reproductive technology (ART), underwent 27 IVF cycles. Analysis of the outcome of IVF including implantation and ongoing pregnancy rates was performed. The data were compared with results from a group of 20 infertile patients with idiopathic infertility undergoing 27 IVF cycles at MGH during the same period. The patients in the two groups were matched for age, basal day 3 levels of follicle stimulating hormone and serum estradiol, and the number and quality of embryos transferred. RESULTS: The response to controlled ovarian hyperstimulation was comparable in the two groups. Significantly lower implantation and ongoing pregnancy rates following IVF and embryo transfer were seen in the utero DES-exposed group compared to the control patients. CONCLUSIONS: Infertile patients with a history of in utero exposure to DES exhibit a significantly impaired implantation rate following IVF, and the outcome of ART remains poor.

Adult↗

PubMed1998

Parenthood motives in IVF-mothers.

This is an exploratory study to answer two questions: (1) does in-vitro fertilization (IVF)-mothers' motivation for parenthood differ from the naturally conceiving mothers' motivation for parenthood? and (2) are IVF-mothers' parenthood motives correlated with their adjustment to parenthood (in the same way as they are for naturally conceiving mothers)? The study sample consisted of 31 mothers with a 24-30 month old, single-born child conceived by homologous IVF and a comparison group of 31 mothers with a naturally conceived child. Self-rating questionnaires were used. Overall, the parenthood motivation pattern was quite similar for IVF-mothers and mothers by natural conception. However, 'identity', 'motherhood' and 'social control' were significantly more important motives for IVF-mothers than for the naturally conceiving mothers. The two former motives remained more important for IVF-mothers when findings were adjusted for the influence of the mothers' age and educational level. Possible explanations for these findings refer to the infertility history. Overall, few correlations between parenthood motives and the adjustment to parenthood were significant, both for the IVF-mothers and for the mothers by natural conception. Further (hypothesis-testing) research is needed to support these findings.

Adaptation, Psychological↗

PubMed1998

GnRH agonists and in vitro fertilization.

GnRH agonists (GnRH-a) were introduced into in vitro fertilization (IVF) superovulation regimens in the late 1980s and have become established as a component of standard regimens in most centers worldwide. Each style of GnRH-a regimen (long, short or ultrashort) is associated with particular advantages and disadvantages. In general the "long protocol" approach tends to be the most widely used. The pros and cons of these different approaches are discussed, as are issues concerned with the management and safety of GnRH-a regimens in IVF.

Female↗

PubMed1995

Color-Doppler velocimetry of uterine arteries in pregnant and nonpregnant patients during multiovulation induction for IVF.

OBJECTIVES: To evaluate uterine artery resistance during multiovulation induction in relation to the implantation rate in patients attending in vitro fertilization (IVF) cycles. PATIENTS: Multiovulation induction for IVF was monitored by daily determination of the pulsatility index (PI) of the uterine arteries, obtained by a transvaginal probe (6.5 MHz) implemented with color-flow imaging. Doppler data were obtained from 5 days before hCG administration to the day of follicular aspiration. One IVF cycle was monitored in 70 patients. In 17 patients, 41 IVF cycles were monitored until a successful attempt occurred. RESULTS: In the 70 patients studied during one IVF attempt, the PI of the uterine arteries significantly varied (P < 0.001) in the different phases of the cycle. In the 24 patients who conceived, a significantly lower PI (P < 0.03) was found throughout the cycle. This result was mainly due to a highly significant difference of PI values observed the day after hCG administration (P < 0.005). In the 17 patients who conceived after 1 to 4 negative in vitro fertilizations, no significant difference in PI was observed in the uterine artery resistance in cycles in which implantation was or was not successful. CONCLUSIONS: Uterine artery resistance varies significantly during phases of the induction therapy. Uterine artery resistance is lower throughout the course of multiovulation induction in patients with higher pregnancy rates. The PI on the day after hCG administration was the best index of pregnancy rate. Low uterine artery resistance was present even in negative attempts in patients who eventually achieved a successful implantation. PI values < or = 3 can be considered a favorable prognostic factor for future IVF cycles.

Adult↗

PubMed1997

The efficacy of trypsin for disinfection of in vitro fertilized bovine embryos exposed to bovine herpesvirus 1.

Bovine cumulus-oocyte complexes (COC) or in vitro fertilized (IVF) embryos were exposed to bovine herpesvirus 1 (BHV-1) during in vitro maturation or co-culture with uterine tubal cells, respectively. Trypsin, at a concentration of 0.25%, was applied (for approximately 90 s) to disinfect either COC or cumulus-free oocytes (CFO) 18 h after insemination, or on day 7 to embryos resulting from infected oocytes. In total, virus was not detected in 71% of 93 samples containing 233 embryos exposed to BHV-1 and trypsin treatment. BHV-1 was detected in 14% and 54% of samples containing a single embryo and five embryos, respectively. In corresponding groups of embryos exposed to BHV-1, then washed but not treated with trypsin (70 samples), 85% and 96% of samples containing one embryo and pooled embryos, respectively, were positive for the virus. There was no effect of trypsin treatment on the development of IVF-embryos. It is concluded that IVF-generated embryos have a greater tendency to carry BHV-1 after experimental exposure to the virus than IVF uterine stage embryos, and that they are more difficult to disinfect by means of the standard trypsin treatment use.

Animals↗

PubMed2006

Aggregating embryonic but not somatic nuclear transfer embryos increases cloning efficiency in cattle.

Our objectives were to compare the cellular and molecular effects of aggregating bovine embryonic vs. somatic cell nuclear transfer (ECNT vs. SCNT) embryos and to determine whether aggregation can improve cattle cloning efficiency. We reconstructed cloned embryos from: 1) morula-derived blastomeres, 2) six adult male ear skin fibroblast lines, 3) one fetal female lung fibroblast line (BFF), and 4) two transgenic clonal strains derived from BFF. Embryos were cultured either singularly (1X) or as aggregates of three (3X). In vitro-fertilized (IVF) 1X and 3X embryos served as controls. After aggregation, the in vitro development of ECNT but not that of SCNT or IVF embryos was strongly compromised. The inner cell mass (ICM), total cell (TC) numbers, and ICM:TC ratios significantly increased for all the aggregates. The relative concentration of the key embryonic transcript POU5F1 (or OCT4) did not correlate with these increases, remaining unchanged in the ECNT and IVF aggregates and decreasing significantly in the SCNT aggregates. Overall, the IVF and 3X ECNT but not the 1X ECNT embryos had significantly higher relative POU5F1 levels than the SCNT embryos. High POU5F1 levels correlated with high in vivo survival, while no such correlation was noted for the ICM:TC ratios. Development to weaning was more than doubled in the ECNT aggregates (10/51 or 20% vs. 7/85 or 8% for 3X vs. 1X, respectively; P < 0.05). In contrast, the SCNT and IVF controls showed no improvement in survival. These data reveal striking biological differences between embryonic and somatic clones in response to aggregation.

Animals↗

PubMed2004

Anti-hyaluronidase oligosaccharide derived from chondroitin sulfate a effectively reduces polyspermy during in vitro fertilization of porcine oocytes.

The present study was conducted to examine the effects of chondroitin sulfate A-derived oligosaccharide (ChSAO) on hyaluronidase activity and in vitro fertilization (IVF) parameters. The activity of hyaluronidase extracted from preincubated boar sperm was completely blocked by ChSAO at concentrations of 10 microg/ml or higher. After in vitro maturation of porcine cumulus-oocyte complexes, some oocytes were freed from their cumulus cells, and cumulus-intact or cumulus-free oocytes were inseminated with sperm in IVF medium containing various concentrations of ChSAO (0.1-100 microg/ml). In cumulus-intact oocytes, the penetration and the polyspermy rates (39% and 28%, respectively) were significantly decreased by treatment with 100 microg/ml ChSAO compared with those of oocytes treated without ChSAO (63% and 52%, respectively). On the contrary, in cumulus-free oocytes, the addition of 10-100 microg/ml ChSAO significantly reduced the polyspermy rate compared with the control (25-30% versus 53%, respectively), whereas ChSAO had no effect on sperm penetration. Interestingly, ChSAO added to IVF medium significantly decreased the number of sperm bound to the zona pellucida (ZP) of cumulus-free oocytes in a concentration-dependent manner between 0.1 and 100 microg/ml. However, ChSAO had no effect on the time course change in ZP modification after oocyte activation by electrostimulation and the incidence of the acrosome-reacted sperm. Treatment with 100 microg/ml ChSAO during IVF of cumulus-free oocytes significantly increased the proportion of development to the blastocyst stage after in vitro insemination. Therefore, the present findings indicate that hyaluronidase-inhibitory ChSAO is an efficient probe for promoting normal fertilization process in terms of an effective decrease in the incidence of polyspermy during IVF of porcine oocytes.

Acrosome Reaction↗