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PubMed1995

A zona biochemical change and spontaneous cortical granule loss in eggs that fail to fertilize in in vitro fertilization.

OBJECTIVE: To determine the extent of biochemical changes in the zona pellucida (ZP) and cortical granule release in eggs failing to fertilize in IVF. DESIGN: After insemination, unactivated eggs without two pronuclei (PN) were studied by high resolution microscopy and fluorescent probes to determine cortical granule density and meiotic stage. After ZP isolation, proteins from individual ZPs were biotinylated, electrophoresed, and visualized by western blots with avidin-chemiluminescence. Controls included mouse unfertilized and fertilized eggs and human germinal vesicle stage oocytes and > or = 3PN eggs. SETTING: University medical center and hospital tertiary care IVF-ET program. RESULTS: Many of the unfertilized eggs were in metaphase and had relatively low cortical granule densities indicative of cortical granule loss. Approximately one half of the ZPs showed evidence of biochemical hardening with a modification of a 90 to 100 x 10(3) molecular (weight) ratio (M(r)) ZP protein. The 3PN eggs had few cortical granules and their ZPs had a pronounced 90 to 100 x 10(3) M(r) modification that was not detected in germinal vesicle stage ZP controls. CONCLUSION: The observed changes in ZP biochemistry and cortical granule quantitation demonstrate that failed fertilization is frequently associated with spontaneous cytoplasmic, but not nuclear, activation. In affected eggs, these changes could prevent fertilization in routine IVF and in cases of reinsemination. The relationship of changes in the ZP and cortical granules to infertility and in vitro culture requires further investigation. The 90 to 100 x 10(3) M(r) ZP protein modification can be detected biochemically with 1/10 of a human ZP.

Animals↗
PubMed1987

Use of individual human follicles to compare oocyte in vitro fertilization to granulosa cell in vitro luteinization.

Granulosa-lutein (G-L) cells from individual follicles aspirated during cycles of in vitro fertilization-embryo transfer were examined after 3 and 6 days in culture. G-L cells from follicles that contained an oocyte that fertilized in vitro were compared with G-L cells from follicles that contained an oocyte that did not fertilize in vitro. Spent culture media was assayed for progesterone at days 3 and 6 of culture and luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor content of G-L cells was determined at day 6. G-L cell cultures from follicles that contained an oocyte that fertilized in vitro produced significantly more progesterone over 3 and 6 days of culture than those obtained from follicles in which the oocyte did not fertilize. Furthermore, LH/hCG receptor content after 6 days was significantly higher in G-L cells obtained from follicles with fertilized oocytes compared with follicles with unfertilized oocytes. Increased progesterone output and LH/hCG receptor acquisition demonstrate more maturation or "luteinization" by G-L cells aspirated from individual follicles that contain oocytes that fertilized in vitro.

Cells, Cultured↗
PubMed1996

In vitro fertilization of in vitro matured pig oocytes: effects of boar and ejaculate fraction.

Ejaculates from 3 young boars were collected on 4 occasions as a series of separate 15-ml fractions. The contribution of different fractions of these ejaculates to observed variability in the quality of the semen when used for IVF was then determined. On the basis of sperm concentration, 3 fractions representing the first peak concentration (Fraction 1), the lowest sperm concentration after Fraction 1 (Fraction 2), and the second peak concentration (Fraction 3) were selected for analysis in vitro. Oocyte-cumulus-granulosa cell complexes were obtained by dissection from slaughterhouse ovaries. In vitro matured oocytes were randomly assigned for fertilization by the 3 semen samples from each boar. Sperm concentration was the same in all the samples during prefertilization incubation, while the final concentration for fertilization was 5 x 10(5) sperm/ml. Data were analysed using ANOVA for a split-plot design. In the presence of fraction effects, Student-Newman-Keuls (SNK) test was used for multiple comparison of treatment means. Oocyte penetration rates differed among fractions (P = 0.001) and varied from 69 to 100% (mean 95.7%) for Fraction 1, from 0 to 100% (mean 53.3%) for Fraction 2, and from 50to 100% (mean 89.9%) for Fraction 3. There were also differences in male pronuclear formation rate (P = 0.028; mean 27.6, 9.3 and 16.4% for Fractions 1, 2 and 3, respectively); in the rate of polyspermy (P = 0.0001; mean 92.3, 31.9 and 76.3% for Fractions 1, 2 and 3, respectively); and in the number of penetrated spermatozoa per oocyte P = 0.002; mean 5.58, 1.94 and 4.07 for Fractions 1, 2, and 3, respectively). The first peak concentration of semen (Fraction 1) showed superiority in fertilizing ability and less variability in penetration rate from replicate to replicate compared with the other 2 fractions. By multiple comparison, Boar 1 showed higher rates of penetration (P < 0.05), male pronuclear formation (P < 0.05) and polyspermy (P < 0.05) than the other 2 boars. There was no fraction-by-boar interaction. The IVM-IVF system adopted proved to be a promising method for boar semen evaluation.

Journal Article↗
PubMedDate not supplied

Human in vitro fertilization.

Human in vitro fertilization has become a widely accepted method of treatment for infertile couples. The list of indications for this technique is rapidly growing and includes oligospermia, endometriosis, and unexplained infertility. The techniques of follicular recruitment, ovum maturation, and follicular aspiration, as well as laboratory culture techniques, are described. The various factors to be taken into consideration when trying to assess possible success of in vitro fertilization and embryo transfer are discussed in this paper.

Chorionic Gonadotropin↗
PubMed2002

In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer.

Three experiments were conducted to evaluate the effect of oocyte and sperm treatments on rates of in vitro fertilization (IVF) in the horse and to determine the capacity of in vitro-matured horse oocytes to be fertilized in vivo. There was no effect of duration of oocyte maturation (24 vs. 42 h) or calcium ionophore concentration during sperm capacitation (3 microM vs. 7.14 microM) on in vitro fertilization rates. Oocytes matured in 100% follicular fluid had significantly higher fertilization (13% to 24%) than did oocytes matured in maturation medium or in 20% follicular fluid (0% to 12%; P < 0.05). There was no significant difference in fertilization rate among 3 sperm treatments utilizing 7.14 microM calcium ionophore (12% to 21%). Of in vitro-matured oocytes recovered 40-44 h after transfer to the oviducts of inseminated mares, 77% showed normal fertilization (2 pronuclei to normal cleavage). Cleavage to 2 or more cells was seen in 22% of oocytes matured in follicular fluid and 63% of oocytes matured in maturation medium; this difference was significant (P < 0.05). We conclude that in vitro-matured horse oocytes are capable of being fertilized at high rates in the appropriate environment and that in vitro maturation of oocytes in follicular fluid increases fertilization rate in vitro but reduces embryo development after fertilization in vivo. Further work is needed to determine the optimum environment for sperm capacitation and IVF in the horse.

Animals↗
PubMed1988

Pregnancies after co-culture of cumulus cells with bovine embryos derived from in-vitro fertilization of in-vitro matured follicular oocytes.

Bovine follicular oocytes surrounded by cumulus cells for more than one-third of their surface were matured, fertilized and developed in vitro utilizing a co-culture system with bovine cumulus cells. Embryos developed into blastocysts were non-surgically transferred to the uteri of cows at Day 6, 7 or 8 (Day 0 = oestrus). Out of 6 recipient cows (19 blastocysts transferred), 3 became pregnant. One of the 3 pregnant cows carried twins. The results of this study demonstrated the viability of embryos obtained from in-vitro maturation of bovine oocytes followed by in-vitro fertilization and culture to the blastocyst stage in vitro.

Animals↗
PubMed2002

Early history of in vitro fertilization.

Although in vitro fertilization (IVF) is used widely for a variety of purposes, it is often not appreciated how this technology was developed. A large number of experiments beginning in 1878 contributed to the first successful reports of IVF over 75 years later. The discovery of sperm capacitation in 1951 was central to the development of IVF technology, and it was rapidly followed by the first convincing reports of IVF in several species. The ability to fertilize oocytes in vitro has allowed major advances to be made into understanding the mechanisms involved in fertilization and early development, and IVF now supports reproductive biotechnology in animals and in humans. This article is a historical review of key experiments that helped to provide the basis for present day IVF procedures, placed into context with current practice.

Acrosome Reaction↗
PubMed1998

Previous fertilization failure with conventional in vitro fertilization is associated with poor outcome of intracytoplasmic sperm injection.

OBJECTIVE: To evaluate the outcome of intracytoplasmic sperm injection (ICSI) in patients with a history of fertilization failure with conventional IVF. DESIGN: Retrospective analysis of 2 years of clinical experience with ICSI. SETTING: Clinical IVF-ET program in a tertiary care referral center. PATIENT(S): The results of ICSI performed between January 1995 and December 1996 were compared between patients with normal semen parameters and a history of fertilization failure (< 20% of oocytes) with conventional IVF and patients with male factor infertility. INTERVENTION(S): In vitro fertilization with ICSI. MAIN OUTCOME MEASURE(S): Parameters examined included oocyte survival, fertilization, embryo cleavage, implantation rates, and clinical pregnancy rates. RESULT(S): Fertilization was achieved with ICSI for all patients during the study period. Although oocyte survival and fertilization did not differ between groups, the pregnancy and implantation rates for patients with a history of idiopathic fertilization failure (20% and 6%, respectively) were significantly lower than those for other patients undergoing ICSI (47% and 22%, respectively). CONCLUSION(S): The outcome of ICSI varied depending on the indication for treatment. Patients who had a history of failed or poor fertilization in vitro with apparently normal semen parameters had significantly lower pregnancy and implantation rates than did patients with either obstructive azoospermia or impaired semen quality.

Adult↗
PubMed1987

Interactions of aged gametes: in vitro fertilization using in vitro-aged sperm and in vivo-aged ova in the mouse.

A study of varying combinations of in vitro-aged sperm and in vivo-aged ova at 3 hr intervals from 0-24 hr resulted in failures at different steps of the fertilization process during in vitro fertilization of mouse ova. Significant decreases caused by sperm aging, ova aging, and sperm X ova aging interaction were found in sperm penetration. Pronuclear formation was not affected by sperm aging and was enhanced by ova aging, and there was a significant effect of sperm X ova aging interaction. Sperm aging significantly influenced the prometaphase stage of the fertilization process. Therefore, it is suggested that the detrimental fertilization effects resulting from aging gametes are due to different mechanisms in sperm and ova, that these mechanisms are affected at different times, and that they affect different steps in the fertilization process.

Aging↗
PubMed1989

In vitro fertilization and development of in vitro matured bovine follicular oocytes.

Bovine follicular oocytes matured in vitro were fertilized in vitro using epididymal spermatozoa from five different bulls and then cultured to the blastocyst stage in vitro. The fertilization rate, based on one pair of pronuclei and presence of one sperm tail, ranged from 55.2 to 64.3%. Embryo development (cleavage to blastocyst stage) ranged from 21.4 to 31.0% of the cultured ova reaching 8 cells at 3 to 4 d after insemination to 1.3 to 3.7% reaching hatched blastocysts at 9 to 10 d. It is concluded that individual variation among bulls is not a significant factor in fertilization and development rates of bovine follicular oocytes when epididymal spermatozoa are used.

Animals↗
PubMed1987

Results of planned in-vitro fertilization programming through the pre-administration of the oestrogen-progesterone combined pill.

The use of an oestrogen-progesterone combined pill permits the induction of ovulation in the absence of any developing follicle. Two treatments were compared. In the first, patients received no prior treatment before stimulation. In the second, combined oestrogen-progesterone treatment was given during approximately two menstrual cycles prior to stimulation. No differences between the two groups were found in relation to oocyte maturity, fertilization in vitro, cleavage, replacement and pregnancy. Fewer luteinizing hormone surges occurred in patients pre-treated with steroids. The utilization of the oestrogen-progesterone combined pill prior to induction of ovulation facilitates the forward planning of patients for in-vitro fertilization.

Clomiphene↗
PubMed1988

The chromosomal normality of in vitro-fertilized rabbit oocytes.

The chromosomal normality of rabbit oocytes fertilized in vitro was examined. Ovum donors were superovulated with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG). Follicular oocytes were collected laparoscopically 9-10 h after hCG treatment and incubated in vitro with spermatozoa capacitated in vivo. Of 267 aspirated oocytes, 191 (71.4%) were fertilized in vitro and developed to the 2- to 8-cell stage 24-48 h after insemination. In the chromosomal studies, 121 (63.4%) were examined. Of these, 94 (77.7%) had a normal diploid complement of chromosomes (2n = 44) and 6 (5%) showed aneuploidy. Of the remaining 21, 20 were triploid and one was tetraploid. The incidence of triploid oocytes after in vitro fertilization was higher than the rate in vivo (16.5% vs. 9.0%, respectively). These triploid oocytes were suspected to be the result of polyspermic fertilization in vitro. In addition, at the Metaphase II stage, 62 (89.9%) of 69 induced, preovulatory oocytes had a normal number of chromosomes.

Animals↗
PubMedDate not supplied

Preeclampsia and eclampsia in parturients from the in vitro fertilization program.

Women from in vitro fertilization program are at higher risk for developing pregnancy induced hypertension, so we analyzed the incidence of preeclampsia and eclampsia in women who conceived in an in vitro fertilization program. A seven-year study included 144 parturients from in vitro fertilization program. The control group consisted of 39,112 parturients from general population. We analyzed only women with singleton pregnancies. There were 2.08% parturients from the in vitro fertilization program, and 0.40% from the control group, with diagnosis of preeclampsia, with chi2=6.24; p<0.05. Eclampsia occurred in 0.69% women who conceived using in vitro fertilization, and in 0.06% parturients from the control group (chi2=2.05; p>0.10). Parturients from in vitro fertilization program are at statistically significantly higher risk for preeclampsia, but not for eclampsia, when compared with women from general population.

Eclampsia↗
PubMed1994

The cost of a successful delivery with in vitro fertilization.

BACKGROUND: The use of in vitro fertilization has engendered considerable debate about who should have the procedure, whether health insurance should cover the cost, and if so, to what extent. We investigated the cost of a successful delivery with in vitro fertilization. METHODS: We calculated the cost per successful delivery with in vitro fertilization (defined as at least one live birth) for a general population of couples undergoing in vitro fertilization and for two subgroups: couples with a diagnosis of tubal disease (who have a better chance of success), and couples in which the woman is over the age of 40 years and the man has a low sperm count (who have a lower chance of success). Information on charges per cycle of in vitro fertilization was obtained from six facilities across the country; delivery rates with this procedure were estimated from the literature. RESULTS: On average, the cost incurred per successful delivery with in vitro fertilization increases from $66,667 for the first cycle of in vitro fertilization to $114,286 by the sixth cycle. The cost increases because with each cycle in which fertilization fails, the probability that a subsequent effort will be successful declines. Sensitivity analyses indicated that the cost per delivery ranges from $44,000 to $211,940. For couples with a better chance of successful in vitro fertilization (i.e., those with a diagnosis of tubal disease), it costs $50,000 per delivery for the first cycle and $72,727 for the sixth. For couples in which the woman is older and there is a diagnosis of male-factor infertility, the cost rises from $160,000 for the first cycle to $800,000 for the sixth. CONCLUSIONS: The debate about insurance coverage for in vitro fertilization must take into account ethical judgments and social values. But analyses of costs and cost effectiveness help elucidate the economic implications of using in vitro fertilization and thus inform the policy discussion.

Adult↗
PubMed2003

Fertilization abnormalities following human in vitro fertilization and intracytoplasmic sperm injection.

Fertilization abnormalities are commonly encountered following in vitro fertilization. The widespread introduction of assisted fertilization methods has rapidly led to a changes in both the incidence and types of these aberrations. Such abnormalities can be identified by careful morphological evaluation at the early zygote stage, of pronuclei as well as of polar body formation, and may be confirmed by cytogenetic assessment. The recognition and understanding of fertilization abnormalities have led to the development of novel techniques aimed at preventing them, as well as recent bold attempts at correction. Removal of one pronucleus may allow some triploid embryos to revert to a normal diploid stage. These new developments should provide insight into the understanding of parthenogenesis, androgenesis, and gynogenesis in the human. Microsc. Res. Tech. 61:358-361, 2003.

Chromosome Aberrations↗
PubMed1992

Pilot study with 120 mg Andriol treatment for couples with a low fertilization rate during in-vitro fertilization.

Treatment with 120 mg/day of Andriol (testosterone undecanoate; Organon, The Netherlands) was given to 11 men whose semen had either failed to fertilize, or had resulted in a less than 33% fertilization rate in a first in-vitro fertilization (IVF) trial. Repeat IVF at the end of a 3 month treatment period resulted in a highly significant increase in the number of oocytes fertilized from 4/95 (4.2%) before to 23/87 (26.4%, P less than 0.001) after Andriol treatment. One couple attained spontaneous conception during the second month of Andriol intake and three pregnancies occurred among the remaining 10 cases undergoing repeat IVF, for a total ongoing pregnancy rate of 36.4%. Sperm concentration but not motility increased in the native semen after Andriol intake, but there were no significant changes in sperm characteristics after preparation. The improvement in pregnancy rate contrasts favourably with the results obtained by repeat IVF of untreated historical controls. The result obtained in the present pilot study should encourage the performance of a larger, placebo-controlled investigation protocol.

Female↗
PubMed1989

Incidence of chromosomal anomalies in early bovine embryos derived from in vitro fertilization.

The incidence of chromosomal anomalies in early bovine embryos derived from follicular oocytes fertilized in vitro using sperm separated by Percoll density gradient centrifugation was investigated. Overall, chromosomal anomalies were observed in 13.7% (138/1005) of embryos. There were 14 haploids (1.4%), 2 hypodiploids (0.2%), 6 hyperdiploids (0.6%), 101 triploids (10.0%), 12 tetraploids (1.2%), 2 diploid/triploid mosaics (0.2%), and 1 diploid/tetraploid mosaic (0.1%). The frequency of triploidy was caused mainly by polyspermy. There was a significant difference in the frequency of embryos with abnormal chromosomes between the two bulls used (P less than 0.005), but Percoll centrifugation did not affect the observed incidence of anomalies. The frequency of chromosomal anomalies in embryos at each stage increased with delay or arrest of development. These results suggest that the incidence of chromosomal anomalies depended on the conditions of in vitro fertilization and the arrest of development.

Aneuploidy↗