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Propofol anaesthesia for ultrasound guided oocyte retrieval: accumulation of the anaesthetic agent in follicular fluid.

Propofol (2,6-diisopropylphenol, Diprivan, ICI-Pharmaceuticals, Manchester, UK) is widely used either as an adjunct in general anaesthesia or as sole anaesthetic agent by the continuous intravenous route and intermittent bolus injections for minor surgical interventions. For several years, we have been using this kind of anaesthesia in transvaginal oocyte retrieval for in-vitro fertilization (IVF), allowing a completely painless puncture on an out-patient basis. From in-vitro studies on mouse oocytes, it appeared that propofol could be deleterious for fertilization in a dose- and time-dependent manner. We therefore investigated the concentrations of propofol in follicular fluid during oocyte retrieval in women. We measured propofol levels in serum and follicular fluid of nine patients at fixed intervals during ultrasound guided oocyte retrieval. Serum levels fluctuated randomly, due to interference from top-off doses of propofol. In follicular fluid, however, we found a steady increase of propofol levels, which was proportional to the total dose of propofol administered. These data indicate that propofol accumulates in follicular fluid. Although it seems unlikely that propofol as used in the present protocol exerts a clinically significant unfavourable effect on IVF, we suggest that the oocyte retrieval procedure should be kept as short as possible, in order to limit the accumulation of the anaesthetic in follicular fluid.

Dose-Response Relationship, Drug↗

Comparison between steroid hormones and cortisol in serum and follicular fluid in stimulated and unstimulated cycles of in vitro fertilization patients.

The present study was undertaken to evaluate the effects of the concentrations of serum and follicular fluid steroids and cortisol levels on the establishment of pregnancies in in vitro fertilization (IVF). Our study group consisted of 42 women (group A) who received gonadotropins for induction of ovulation for IVF. The control group included 23 women (group B) who underwent in vitro fertilization without stimulation. Serum estradiol and progesterone levels were significantly higher in group A than in group B. Serum and follicular fluid cortisol levels were similar in both groups A and B. There was no significant difference in the fertilization rates of the stimulated or unstimulated cycles. However, there were no pregnancies in group B whereas there was a 28.5% pregnancy rate in group A. There were no correlations between the estradiol, progesterone and cortisol levels when compared to the oocyte maturity and the fertilization rates in both groups of patients.

Adult↗

Chromosome investigations in early life. I. Human oocytes recovered in an IVF programme.

Fifty-five oocytes recovered in an in-vitro fertilization (IVF) programme and remaining unfertilized when observed 42 h after insemination were prepared for chromosomal analysis. Sixteen oocytes displayed no polar body at the time of fixation and were supposed to be in metaphase I. In fact only two of them were in diakinesis, the others containing a diploid set of metaphase II chromosomes indicating that in 89% of the cases oocytes achieved meiosis without any extrusion of the first polar body. Thirty-nine oocytes in metaphase II were analysed. Nine were abnormal showing three D nullosomies, one G disomy, one double disomy for a 3 and a D chromosome, one deletion of the long arm of a G chromosome, one cell with extra chromosomes and/or chromosome breaks, one endoreduplication and one tetraploidy. The overall rate of abnormalities reached 22%. This high rate of chromosome anomalies can be explained by the nature of this population of fertilization failure, the frequently advanced maternal age and the use of superovulation treatments.

Chromosome Aberrations↗

Filariasis infection is a probable cause of implantation failure in in vitro fertilization cycles.

OBJECTIVE: To describe a parasitic infection that probably affected the implantation of good-quality embryos in an in vitro fertilization (IVF) cycle. DESIGN: Case report. SETTING: Tertiary care center in a university hospital. PATIENT(S): A 36-year-old Caucasian female with primary unexplained infertility. The patient underwent two cycles of IVF with good-quality embryos transferred; however, no pregnancy ensued despite adequate luteal support. INTERVENTION(S): In vitro fertilization cycles, CBC, blood smear, evaluation for eosinophilia including serological evaluation for parasitic infections. MAIN OUTCOME MEASURE(S): Pregnancy. RESULT(S): Following treatment for filariasis, a repeat IVF cycle using the same stimulation protocol yielded a full-term pregnancy. CONCLUSION(S): This case is of particular importance because, to our knowledge, it is the first to describe a parasitic infection that probably affected the implantation of good-quality embryos in IVF cycles.

Adult↗

Effect of glycosaminoglycans on the preimplantation development of embryos derived from in vitro fertilization and somatic cell nuclear transfer.

The purpose of this study was to evaluate the effect of glycosaminoglycans (GAGs) added to the culture medium on the developmental competence of bovine embryos derived from in vitro fertilization (IVF) and from somatic cell nuclear transfer (SCNT). In vitro-matured oocytes were either inseminated with 1 x 10(6) spermatozoa mL(-1) or enucleated and reconstructed with bovine adult ear fibroblasts by SCNT. The embryos were then cultured in modified synthetic oviduct fluid (mSOF) containing 8 mg mL(-1) bovine serum albumin (BSA) (control mSOF) or control mSOF supplemented with various GAGs (hyaluronic acid, heparin or chondroitin sulfate) in a dose-dependent manner (0.1, 0.5 or 1.0 mg mL(-1)). Developmental competence was evaluated by monitoring the numbers of 2-cell embryos, 8-16-cell embryos and blastocysts. The mean cell number of flattened blastocysts stained with 5 microM bisbenzimide on Day 8 was counted. The percentage of blastocyst formation (IVF and SCNT embryos) from cleaved embryos was significantly higher (P < 0.05) in control mSOF supplemented with 0.5 mg mL(-1) hyaluronic acid (45% and 47%), heparin (40% and 47%) or chondroitin sulfate (38% and 44%) compared with control mSOF (30-31% and 30-33%). When compared with the efficacy of 0.5 mg mL(-1) GAGs, no significant differences were observed in the developmental competence of both IVF and SCNT embryos. Supplementing control mSOF with 0.5 mg mL(-1) GAGs had no effect on the cell number of IVF embryos. In contrast, supplementing 0.5 mg mL(-1) of hyaluronic acid, heparin or chondroitin sulfate to control mSOF significantly (P < 0.05) increased the numbers of total cells (93-98 v. 88 cells) and trophectoderm (TE) cells (64-66 v. 55 cells), and decreased the inner cell mass (ICM) to TE cell ratio (48.2-49.8 v. 61.3) in SCNT blastocysts compared with embryos in control mSOF. In conclusion, supplementation of culture media with GAGs may improve the development of bovine IVM-IVF and SCNT embryos to the blastocyst stage. The GAGs increased the quality of blastocysts by increasing total cell numbers in the SCNT embryos.

Animals↗

In vitro development to blastocysts of early porcine embryos produced in vivo or in vitro.

The objective of this study was to compare the development of porcine embryos from the 2- and 4-cell stages to the blastocyst stage after in vivo or in vitro fertilization and in vivo or in vitro culture. Early-stage embryos were collected either from superovulated gilts 36 h after the second mating or after in vitro fertilization (IVF) of in vivo-matured oocytes, both followed by in vitro culture to the blastocyst stage. Blastocysts collected from superovulated donors served as controls. In the first experiment, a total of 821 2- and 4-cell embryos derived from in vivo-fertilized oocytes was cultured either in medium NCSU 23, modified Whittens' medium or modified KRB for 5 d. Significantly (P < 0.05 and P < 0.001) more embryos overcame the 4-cell block and developed to the blastocyst stage in medium NCSU 23 than in the 2 other culture media. Hatching was only observed in medium NCSU 23. In the second experiment, embryos derived from in vivo-matured oocytes fertilized in vitro were cultured in medium NCSU 23. Of 1869 mature oocytes 781 (41.8%) cleaved within 48 h after in vitro fertilization. A total of 715 embryos was cultured to the morula and blastocyst stages, and 410 (57.3%) overcame the developmental block stage, with 358 embryos (50.1%) developing to the morula and blastocyst stages. None of the embryos hatched, and the number of nuclei was significantly (P < 0.05) lower compared with that of in vivo-fertilized embryos (18.9 +/- 9.8 vs 31.2 +/- 5.8). In the third experiment, 156 blastocysts derived from in vitro fertilization and 276 blastocysts derived from in vivo fertilization and in vitro culture were transferred into synchronized recipients, while 164 blastocysts were transferred immediately after collection into 6 recipients, resulting in a pregnancy rate of 83.3%, with 35 piglets (on average 7.0) born. From the in vitro-cultured embryos, 58.3% (7/12) of the recipients remained pregnant at Day 35 after transfer, but only 33.3% maintained pregnancy to term, and 14 piglets (on average 3.5) were born. In contrast, the transfer of embryos derived from in vitro-fertilized oocytes did not result in pregnancies. It is concluded that 1) NCSU 23 is superior to modified Whittens' medium and modified KRB and 2) blastocysts derived from in vitro fertilization have reduced viability as indicated by the lower number of nuclei and failure to induce pregnancy upon transfer into recipients.

Journal Article↗

How does polyspermy happen in mammalian oocytes?

Polyspermy is one of the most commonly observed abnormal types of fertilization in mammalian oocytes. In vitro fertilization (IVF) provides approaches to study the mechanisms by which oocytes block polyspermic fertilization. Accumulated data indicate that oocyte, sperm and insemination conditions are all related to the occurrence of polyspermic fertilization. A high proportion of immature and aged oocytes showed polyspermy as compared with mature oocytes. Preincubation of oocytes and/or sperm with oviductal epithelial cells or collected oviductal fluid before IVF reduces polyspermic penetration. Recently, it was found that an abnormal zona pellucida is one of main causes of polyspermy in human eggs. A high proportion of polyspermy has resulted from the use of a high concentration of capacitated spermatozoa at the site of fertilization, irrespective of in the in vivo or in vitro environment. Oviductal secretions or oviductal epithelial cells themselves can regulate the number of spermatozoa reaching or binding to the zona pellucida thus reducing multiple sperm penetration. Suboptimal in vitro conditions, such as supplementations in IVF media, pH, and temperature during IVF, also induce polyspermic fertilization in some mammals. Species-specific differences are present regarding the relationship between insemination conditions and polyspermy.

Animals↗

Reproductive biology and IVF: ovarian stimulation and luteal phase consequences.

Most clinicians working in in vitro fertilization (IVF) centers worldwide have taken for granted for more than a decade the paradigm of so-called 'controlled' ovarian hyperstimulation, using maximum stimulation by exogenous gonadotropins, together with the gonadotropin-releasing hormone (GnRH) agonist long-protocol. Potential detrimental effects of this approach with regard to oocyte quality, corpus luteum function and endometrial receptivity have been largely ignored. These factors might by themselves have a major impact on IVF outcome and should therefore be considered seriously. The recent introduction of GnRH antagonists along with the current emphasis on the need for transfer of a reduced number of embryos enables a careful re-evaluation of current IVF strategies. We can now render stimulation protocols simpler, starting with a spontaneous menstrual cycle, allowing for more subtle interference with single dominant follicle selection. Here, we discuss recent approaches to ovarian stimulation, the induction of oocyte maturation, and effects of these altered follicular phase interventions on corpus luteum function following ovarian stimulation.

Adult↗

Contextualizing a medical breakthrough: an overview of the case of IVF.

In this article I examine the ways in which the technology of in vitro fertilization (IVF) is embedded in prevailing sociocultural perceptions and power relations in North American and Western European contexts. From this perspective, I look at the efforts involved in developing reproductive technologies and at the particular direction such attempts have taken. I then examine the policies and laws through which IVF is regulated; the technical characteristics of the technology and its implementation; the social implications of IVF for poorer women, women of color, and single and lesbian women; and the media coverage of the subject. Women's responses to IVF and their roles in its shaping are also addressed. I conclude with a reference to the social and cultural repercussions of IVF in the industrialized world. The overview suggests that IVF embodies social continuity while also encapsulating a potential for radical social transformations.

Attitude to Health↗

Fertilization and cleavage rates of heparin-exposed human oocytes in vitro, and the effect of heparin on the acrosome reaction.

Fertilization and cleavage rates were compared in 1024 heparin-exposed and nonexposed human oocytes recovered from 183 consecutive in vitro fertilization (IVF) cycles. Heparinized Ham's F-10 medium, (Gibco, Grand Island, NY) 1.0 ml (2.0 mg heparin/ml) was added to bloody follicular fluid; clear follicular aspirates did not receive heparin. Fertilization and cleavage rates for heparin-exposed (n = 714) and nonexposed (n = 310) oocytes were not significantly different: 63.9 versus 61.6% fertilized (chi 2 = 0.472); 89.3 versus 87.4% of fertilized eggs cleaved (chi 2 = 0.445). A subset of 100 patients, each contributing both heparin-exposed and nonexposed oocytes, also was evaluated. Fertilization and cleavage rates were not significantly different: 59.1 versus 60.8% fertilized (chi 2 = 0.192); 87.6 versus 87.2% of fertilized oocytes cleaved (chi 2 = 0.014). A modified triple stain was used to evaluate the acrosome reaction rate of sperm that had been coincubated with 76 oocytes from ten patients. There was no significant difference in the proportion of viable acrosome-reacted sperm following incubation with heparin-exposed (1.9 +/- 1.0%) versus nonexposed (2.3 +/- 1.2%) (mean +/- standard deviation [SD]) oocytes. The addition of heparin to follicular fluid at the time of oocyte recovery for IVF has no apparent effect on fertilization or cleavage in vitro, nor any influence on the acrosome reaction.

Acrosome↗

The incidence of major clinical complications in a Dutch transport IVF programme.

Four different major clinical complications were identified in a retrospective analysis of 2495 in-vitro fertilization (IVF) cycles resulting in oocyte retrieval. The severe form of ovarian hyperstimulation syndrome (OHSS) occurred in 18 patients, giving a prevalence for this complication of 0.7%. Seven (39%) of these 18 patients had previously been diagnosed as having polycystic ovaries. Eleven patients were admitted with moderate OHSS. Adnexal torsion was diagnosed in two patients. Ovariectomy was considered necessary in both cases. Complications of the transvaginal procedure occurred in seven cases (0.3%): one patient had an acute appendicitis with puncture holes in the appendix, six patients were admitted shortly after oocyte retrieval with a pelvic inflammatory disease. Of the 624 pregnancies obtained, 13 were ectopic, giving an ectopic pregnancy rate of 2.1%. It is concluded that serious clinical complications of IVF treatment are rare. However, patients should be counselled for the occurrence of serious procedure-related complications before entering an IVF programme.

Adnexal Diseases↗

Dimeric inhibins and activin A in human follicular fluid and oocyte-cumulus culture medium.

The concentrations of inhibin A, inhibin B and activin A in follicular fluid and oocyte culture medium were analysed to investigate the production of these peptide hormones by ovarian granulosa cells and oocyte-cumulus complexes, as well as their potential as possible biochemical markers for oocyte quality and fertilizing capacity. Follicular fluids were collected from individual follicles during oocyte retrieval for in-vitro fertilization (IVF). Oocyte-cumulus culture media were collected after in-vitro insemination. The concentrations of dimeric inhibin A, inhibin B and activin A were measured using two-site enzyme-linked immunosorbent assays in the follicular fluid and matched oocyte culture medium. Hormone concentrations were compared with oocyte quality and fertilizing capacity. The concentration of inhibin A in follicular fluid increased while that of inhibin B decreased with increasing follicle size. Follicular fluid concentrations of inhibin A inhibin B and activin A were not significantly different in follicles with differing oocyte quality. Oocyte culture medium concentrations of activin A were significantly higher in morphologically good quality oocytes. There was no relationship between the concentrations of the three hormones and oocyte fertilizing capacity. This study confirms that follicular fluid concentrations of inhibin A may prove to be a marker of follicular growth and maturation. Higher concentrations of activin A produced by good quality oocyte-cumulus complexes suggest that activin A may play a role in oocyte maturation.

Activins↗

Treatment policy after poor fertilization in the first IVF cycle.

PURPOSE: The chance of recurrence of poor fertilization in a second in vitro fertilization (IVF) cycle was assessed. METHODS: Total fertilization failure was defined, and the relationship between the fertilization rate and the number of motile sperm cells per milliliter of semen was assessed. Patients with a total fertilization failure or poor fertilization (20% or less of the oocytes fertilized) were divided into three subgroups with different chances of fertilization and were followed in a subsequent IVF cycle. RESULTS: The recurrence rate of total fertilization failure was high in all three groups (45-70%), and poor fertilization frequently occurred in the second cycle (50-75%). CONCLUSIONS: Poor fertilization frequently recurs in the second IVF cycle. The use of intracytoplasmic sperm injection could be considered after fertilization of 20% or less of oocytes in the first cycle, irrespective of the number of motile sperm cells per milliliter of semen.

Female↗

Influence of body mass index and self-administration of hCG on the outcome of IVF cycles: a prospective cohort study.

Final maturation of the oocyte in in vitro fertilization (IVF) cycles is achieved through the administration of a timed injection of human chorionic gonadotrophin (hCG). The success of mature oocyte retrieval is dependent on serum concentrations of the hormone reaching values capable of initiating meiosis and triggering the release of the cumulus-oocyte complex into the follicular fluid. The objective of this prospective cohort study was to examine the effect of adiposity, as measured by body mass index (BMI), on serum concentrations of hCG and gonadotrophins and to relate this to IVF outcome. A comparison was also made between professionally and non-professionally administered hCG to assess any possible effect on cycle parameters. A total of 50 patients with a high BMI (> or = 26 kg m(-2)) who underwent IVF treatment at the Assisted Conception Unit, St James's University Hospital, Leeds, was recruited prospectively into the study. They were matched with 50 patients with a normal BMI (18-25 kg m(-2)) who acted as a control group. The two groups were matched for age (mean of 32 years and range of 22-42 years) and cause of infertility. Serum gonadotrophins, oestradiol and hCG concentrations, measured at the time of oocyte retrieval, and the clinical outcome of the two groups were compared. Patients with a high BMI had a significantly lower mean serum hCG concentration compared with controls (63.9 versus 99.6 iu l(-1), P < 0.0003). They also required a higher dosage of gonadotrophin (3660 versus 3007 iu) to achieve follicular maturation than the controls. Similarly, the high BMI group of patients had higher serum concentrations of follicle-stimulating hormone (FSH) (12.3 versus 11.2 iu l(-1)) and lower oestradiol (3499 versus 3506 pmol l(-1)) compared with controls. Patients with a high BMI had significantly fewer oocytes aspirated, resulting in a significant decrease in the oocyte:follicle ratio compared with controls (33.9 versus 41.7, P < 0.05). The fertilization rate (46.2 versus 61.3%, P < 0.05) and clinical pregnancy rate per cycle (26.6 versus 37.1%, P < 0.05) were also lower in the patients with high BMI compared with those with normal BMI. The administration of hCG by the patient or her partner did not have a significant effect on clinical outcome. The mean serum hCG at the time of oocyte recovery was equivalent in both groups (87.1 versus 89.7 iu l(-1)). Furthermore, the oocyte:follicle ratio (0.73 versus 0.72), fertilization rate (46.2 versus 54.2%) and clinical pregnancy rate (38.9 versus 36.5%) were similar. These findings indicate that high BMI is detrimental to the success of IVF treatment and has an important influence on the distribution and metabolism of hCG. The results also indicate that non-professional administration of hCG does not compromise cycle outcome.

Adult↗

Perinatal outcomes in singletons following in vitro fertilization: a meta-analysis.

OBJECTIVE: To estimate whether singleton pregnancies following in vitro fertilization (IVF) are at higher risk of perinatal mortality, preterm delivery, small for gestational age, and low or very low birth weight compared with spontaneous conceptions in studies that adjusted for age and parity. DATA SOURCES: We searched MEDLINE, BIOSIS, Doctoral Dissertations On-Line, bibliographies, and conference proceedings for studies from 1978-2002 using the terms "in vitro fertilization," "female infertility therapy," and "reproductive techniques" combined with "fetal death," "mortality," "fetal growth restriction," "small for gestational age," "birth weight," "premature labor," "pre-term delivery," "infant," "obstetric," "perinatal," and "neonatal." METHODS OF STUDY SELECTION: Inclusion criteria were singleton pregnancies following IVF compared with spontaneous conceptions, control for maternal age and parity; 1 of the above outcomes; and risk ratios or data to determine them. Study selection and data abstraction were performed in duplicate after removing identifying information. TABULATION, INTEGRATION, AND RESULTS: Fifteen studies comprising 12,283 IVF and 1.9 million spontaneously conceived singletons were identified. Random-effects meta-analysis was performed. Compared with spontaneous conceptions, IVF singleton pregnancies were associated with significantly higher odds of each of the perinatal outcomes examined: perinatal mortality (odds ratio [OR] 2.2; 95% confidence interval [CI] 1.6, 3.0), preterm delivery (OR 2.0; 95% CI 1.7, 2.2), low birth weight (OR 1.8; 95% CI 1.4, 2.2), very low birth weight (OR 2.7; 95% CI 2.3, 3.1), and small for gestational age (OR 1.6; 95% CI 1.3, 2.0). Statistical heterogeneity was noted only for preterm delivery and low birth weight. Sensitivity analyses revealed no significant changes in results. Early preterm delivery, spontaneous preterm delivery, placenta previa, gestational diabetes, preeclampsia, and neonatal intensive care admission were also significantly more prevalent in the IVF group. CONCLUSION: In vitro fertilization patients should be advised of the increased risk for adverse perinatal outcomes. Obstetricians should not only manage these pregnancies as high risk but also avoid iatrogenic harm caused by elective preterm labor induction or cesarean.

Female↗

Value of the hamster oocyte test and computerised measurements of sperm motility in predicting if four or more viable embryos will be obtained in an IVF cycle.

The experimental group consisted of men from 81 couples waiting for in vitro fertilization (IVF), about half of whom had sperm dysfunction defined by a negative post-coital test. A diagnostic semen sample was subjected to a hamster oocyte penetration test (HOPT) after stimulation of the acrosome reaction with A23187 +/- pentoxifylline and to computerized sperm motility measurements (CASA) as well as conventional semen analysis according to the WHO protocol. Logistic regression was used to identify parameters that predicted the probability of achieving four or more viable embryos at IVF among the 65 couples from whom four or more oocytes were collected. The number of oocytes available and whether the woman had previously been pregnant (ever pregnant) were important factors but once these had been taken into account a number of sperm parameters had additional predictive power. The most useful of these were the percentage sperm static (CASA) or the percent sperm progressively motile (conventional semen analysis) in the Percoll preparation. A model incorporating the number of oocytes collected, ever pregnant and percentage sperm static achieved 85% correct prediction of outcome in the experimental dataset but only 62% correct prediction in an independent set of 280 IVF cycles. The percentage of hamster oocytes penetrated was a significant predictor but had no advantage over simple motility measurements. The results illustrate the difficulty of basing a prognosis for achieving satisfactory fertilization in IVF on the properties of spermatozoa.

Adult↗

A follow-up study of sperm preparation for IVF by swim-up in a solution of hyaluronate.

Spermatozoa prepared for in-vitro fertilization (IVF) by swim-up in a balanced salt solution containing hyaluronate gave rates of fertilization, cleavage and pregnancy which were not significantly different from those obtained with sperm prepared by swim-up in standard IVF medium followed by centrifugation. However, the content of prostaglandin F2alpha in the final sperm suspension was higher using hyaluronate but this seemed to be of no consequence for IVF. Thus, preparation of normal sperm samples for IVF may be simplified by performing swim-up in a balanced salt solution containing hyaluronate.

Cell Separation↗

The selection criteria on an IVF program can remove the association between maternal age and implantation.

BACKGROUND: 1190 consecutive in vitro fertilization (IVF) treatment cycles from the Southampton University/BUPA Chalybeate unit, spanning a four year period, were studied retrospectively in order to assess the relationship between maternal age and implantation. Our aim was to evaluate the hypothesis that the number of transferred embryos can be determined by age alone. METHOD: The cases were allocated to two age groups, Group 1 was composed of patients of less than or equal to 35 years of age and Group 2 of patients greater than 35 years of age. RESULTS: We found that the selection criteria used in our programme for abandoning treatment cycles led to significantly more older patients being excluded from oocyte collection (p < 0.001). The patients from both groups that progressed to oocyte collection and embryo transfer showed no significant difference in embryo implantation. The overall implantation rate (12.4%) and clinical pregnancy rate per embryo transfer (22.8%) were achieved by being able to transfer comparable numbers of embryos in both age groups and in spite of the younger age group having a significantly better quality of transferred embryos. CONCLUSION: Although advancing maternal age predisposes to a reduced chance of success from IVF treatment, maternal age alone was not a useful predictor of embryo implantation or endometrial receptivity in completed IVF treatment cycles.

Adult↗