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PubMed1995

Female aging and superovulation induction for IVF.

The success of in vitro fertilization (IVF) has been reported to be adversely affected by increasing women's age. The main reason being that older women more frequently show a poorer response to stimulation. Information about such variability in ovarian responsiveness to stimulation is important for assessing the chances for successful pregnancy in older women undergoing assisted reproduction, because those with limited ovarian reserve should be counseled to consider other therapeutic options without delay. I present results demonstrating the ovarian response to gonadotropin stimulation and the success rate in 746 IVF cycles. A brief overview on current methods of evaluating ovarian reserve as a means to identify the poor responder, and a discussion on the efficacy of several stimulation regimens for old women with limited ovarian reserve is also provided.

Adult↗
PubMed2004

[Effect of early cleavage of embryos after intracytoplasmic sperm injection (ICSI) into oocytes on results of fertilization in vitro (IVF) and embryo transfer (ET)].

OBJECTIVE: To evaluate the influence of early cleavage in embryos obtained using ICSI on the success rate of IVF-ET. DESIGN: A prospective study. SETTING: Centre of Assisted Reproduction, Department of Obstetrics and Gynecology, Medical Faculty, Palacký University, Olomouc. METHODS: The rate of the first cleavage of embryos was checked between 23 to 26 hours after ICSI in 115 cycles IVF-ET. "Early Cleavage" (EC) embryos which were already cleaved were cultivated separately from the "slow" (No Early Cleavage, NEC) embryos. After 3 days cultivation, no more than three 6- to 8-cell embryos were transferred. Only EC embryos were transferred in 43 cycles and only NEC embryos were transferred also in 43 cycles. The remaining 29 cycles, where both EC and NEC embryos had to be transferred, were not evaluated. In both groups the following entry parameters were compared: age of the patient, number of fertilized oocytes, fertilization rate (FR), number of frozen and evaluated embryos, number of EC embryos recorded and number of transferred embryos. The outcome parameters compared were: pregnancy rate (PR), number of gestational sacs with heart beats per number of transferred embryos (implantation rate, IR), abortions (AB) and the frequency of multiple pregnancy. Statistical evaluation was performed by t-test, chi 2 test and by Sheffe analysis. RESULTS: EC embryos were found in 57% of cycles (n = 66) after ICSI. Early cleavage was observed in 198 of 563 embryos (35%). Among entry parameters the only statistically significant differences (P < 0.05) were found in the number of retrieved oocytes as well as in the ratio of frozen embryos which were higher in the cycles with the transfer of only EC embryos. FR was the same (74%) in both types of transfer. Outcome parameters in the transfer of only EC versus NEC embryos were higher in PR (40% vs. 33%), about equal in IR (27% vs. 25%) and lower in AB (2% vs. 5%). Moreover, multiple pregnancies were less frequent in the transfer of only EC embryos: 65% of singleton pregnancy, 35% of twins and no triplet versus 36%, 57% and 7%, resp., in the transfer of only NEC embryos. The differences in outcome parameters are not statistically significant (P > 0.05) because of small numbers. CONCLUSION: The speed of the first cell cleavage is a useful additional criterion for better evaluation of quality and viability of embryos.

Adult↗
PubMed1994

The willingness to pay for in vitro fertilization: a pilot study using contingent valuation.

The use of in vitro fertilization (IVF) has increased rapidly in recent years and debate has focused on whether health insurance should cover the procedure. We developed and disseminated a contingent valuation survey to investigate how individuals value IVF treatment. Couples pursuing in vitro fertilization can be viewed as purchasing an increased probability of conceiving and bearing a child. In our survey, respondents were asked if they would pay stated amounts for IVF, under various assumptions about the probability of success. The survey explores the ex post perspective, (respondents' willingness to pay (WTP) for IVF in the event that they are infertile), the ex ante perspective, (WTP for IVF insurance, assuming respondents do not know their infertility status), WTP for a public IVF program, and how respondents value IVF relative to mortality risk reduction. Among 150 respondents who were potential childbearers, average WTP was $17,730 for a 10% chance at having a child through IVF in the event of infertility. Average WTP was $865 for a lifetime insurance benefit providing access to a 10% chance. Among 231 respondents of all ages, average WTP was $32 per year in taxes for a public program giving 1,200 couples per year in Massachusetts a 10% chance. The estimated implied ex post WTP per statistical baby was $177,730, while ex ante WTP per statistical baby was $1.8 million. The results of regression analyses were consistent with theoretical predictions, (e.g., increasing WTP with household income). The contingent valuation method is a potentially useful tool in understanding how people value the benefits of IVF. Further research is needed to test the validity of this methodology.

Adult↗
PubMed2001

Embryo transfer after autologous endometrial coculture improves pregnancy rates.

Since the first in-vitro fertilization (IVF) baby born in 1978, the technique of in-vitro fertilization-embryo transfer (IVF-ET) has been well established and widely used by clinicians all over the world. However, the success rate of IVF-ET still remains relatively low. Sub-optimal embryo culture condition may consider as a factor contribute to the poor success rate. Autologous endometrial coculture is a newly developed technique able to enhance development of fertilized eggs using the patient's own endometrial cells. A clinical tryout was conducted in our institute. Patients underwent either classic IVF (n = 82) (i.e., control group) or autologous endometrial coculture (n = 134) (i.e., coculture group). The clinical pregnancy rate (per patient) was significantly higher in coculture group (48.5%) than control group (24.3%). Patients were further subgrouped according to their age. For each subgroup, the clinical pregnancy rates were again consistently higher and the miscarriage rates were consistently lower in the coculture group than in the control groups. Coculture also increased clinical pregnancy rate of patients who failed IVF at least twice from 0% to 26.6%. Our data confirmed that autologous endometrial coculture is an effective method to improve the success rate of IVF-ET. The evidence of improving clinical pregnancy rates and reducing abortion rates after coculture suggest that coculture indeed improved success rates by improving embryo quality.

Adult↗
PubMed1990

Isolation, in vitro maturation, and fertilization of germinal vesicle oocytes obtained from the intact murine ovary.

The purpose of this investigation was to attempt to develop a process, utilizing a murine model, which would allow more efficient harvesting from the intact ovary and maturation in vitro of germinal vesicle (GV) oocytes. The recovery process yielded 25.5 +/- 4.5 (mean +/- SE) cumulus-free GV oocytes per animal. Treatment groups included culture medium (CM) supplemented with either estradiol (E2), follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG), or prolactin (PRL). Among the hormone-free controls 83.2 +/- 1.6% of oocytes underwent GV breakdown, whereas 25.3 +/- 2.6% developed to the first polar body stage (PB-1) following 18 hr of incubation (n = 29 trials). Oocytes progressing to the PB-1 stage were inseminated in vitro. In vitro fertilization (IVF) of pooled in vitro matured (IVM) PB-1 oocytes (judged by two-cell formation) was 19.9%, which was significantly lower than in the group of in vivo matured oocytes (74.4%). E2 significantly increased the percentage of GV breakdown (control, 76.8 +/- 2.5%; E2 at 10 ng/ml, 92.9 +/- 2.5%, P less than 0.001; E2 at 100 ng/ml, 93.7 +/- 2.1%, P less than 0.001; and E2 at 1 micrograms/ml, 86.7 +/- 3.3%, P less than 0.05) but not PB-1 formation. Neither FSH nor hCG significantly increased GV breakdown or PB-1 formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Effects of gonadotropin treatment on ovarian follicle growth, oocyte quality and in vitro fertilization of oocytes in prepubertal gilts.

This study was undertaken to compare the effects of FSH-pituitary (FSH-P), eCG, and a combination of gonadotropins containing 400 IU eCG and 200 IU hCG (PG 600) on the growth of large follicles, oocyte quality and in vitro fertilization (IVF) rate of in vitro matured (IVM) oocytes in prepubertal gilts. The ovaries were removed via midventral laparotomy 48 h (Experiment 1) or 72 h (Experiment 2) after the first injection. In Experiment 1, 30 gilts received 1 of 5 treatments: 1) saline (3 ml i.m., once, n = 6); 2) FSH-P8 (8 mg i.m., twice, with a 24-h interval, n = 6); 3) FSH-P16 (16 mg i.m., twice, with a 24-h interval, n = 6; 4) eCG (1000 IU i.m., once, n = 6); or 5) PG 600 (5 ml i.m., once, n = 6). Compared with saline, treatment with PG 600 or eCG induced significant (P < 0.05) growth of large follicles (> or = 6 mm). In Experiment 2, 16 gilts received 1 of 5 treatments: 1) saline (n = 4); 2) FSH-P8 (n = 4); 3) FSH-P16 (n = 4); 4) eCG (n = 4), or 5) PG 600 (n = 4). The same injection protocol as in Experiment 1 was used. Compared with treatment with FSH-P8 or FSH-P16, eCG increased (P<0.05) the number of large follicles. The proportion of good oocytes was increased (P<0.05) with FSH-P8 or FSH-P16 compared with treatment with eCG or PG 600. Moreover, oocytes from eCG-treated gilts had a greater (P<0.05) rate of male and female pronuclei than FSH-P or saline-treated gilts. In conclusion, treatment with FSH-P resulted in a higher proportion of oocytes with multilayer cumulus cells, whereas treatment with eCG resulted in higher pronuclear rates following in vitro fertilization in prepubertal gilts.

Journal Article↗
PubMed2002

Analysis of differential messenger RNA expression between bovine blastocysts produced in different culture systems: implications for blastocyst quality.

Using reverse transcriptase-amplified fragment length polymorphism (RT-AFLP) analysis of differential mRNA expression and semiquantitative reverse transcriptase-polymerase chain reaction, we compared mRNA expression in bovine blastocysts from 4 sources, known to differ in quality in terms of their ability to withstand cryopreservation: 1) in vitro culture in synthetic oviduct fluid of in vitro-matured (IVM)/in vitro fertilized (IVF) zygotes; 2) in vitro culture in TCM-199 supplemented with granulosa cells (coculture) of IVM/IVF zygotes; 3) in vivo culture in the ewe oviduct of IVM/IVF zygotes; or 4) superovulation, artificial insemination, and nonsurgical embryo recovery. Total mRNA was isolated from pools of blastocysts and reverse transcription was performed. Triplicate reactions from each sample were displayed, and only consistent banding variations were recorded. Using AFLP-differential display assay, we found that cDNA banding patterns are highly conserved between the 4 groups of blastocysts studied; however, there was a difference of 7% in bands either missing or expressed across the groups. Fifty bands were reamplified, and a sequence comparison search revealed similarity of 14 isolated fragments to ribosomal and mitochondrial genes, 16 matched to described cDNA, and 20 corresponded to unknown sequences that may represent novel genes. The study of 7 differentially expressed mRNAs known to be involved in developmental process in the embryo suggests roles for apoptosis, oxidative stress, gap junctions, and differentiation in the determination of embryo quality. The aberrant transcription patterns detected in in vitro-produced bovine embryos compared with those produced in vivo may explain their reduced quality in terms of viability after cryopreservation.

Animals↗
PubMed1994

Effect of growth factors on bovine blastocyst development in a serum-free medium.

To investigate the effect of growth factors on pre-implantation development, bovine zygotes, produced by in vitro fertilization (IVF) of in vitro-matured (IVM) oocytes, were cultured in a serum-free medium to which the following growth factors were added one at a time: epidermal growth factor (EGF), acidic fibroblast growth factor (a-FGF), insulin-like growth factor-II (IGF-II), platelet-derived growth factor from human platelets (PDGF), and platelet-derived growth factor-AB, human, recombinant (PDGF-AB). All growth factors were added at a dose of either 10 or 50 ng/ml, except PDGF which was added at a dose of either 5 or 15 ng/ml. The control medium was TCM 199 supplemented with sodium pyruvate (0.25 mmol/l), BSA (10 mg/ml), insulin (5 micrograms/ml), transferrin (5 micrograms/ml), and sodium selenite (5 ng/ml). Embryos were cultured for 8 days (day of insemination = Day 0). The mean percentages of first cleavage on Day 2 varied from 67% to 86% and the differences between the 2 doses, or between the control and growth factor-treated groups were not significant (p > or = 0.13). The effects of the two doses on subsequent development up to the blastocyst stage did not differ either (p > or = 0.12). There was no stimulatory effect of any of the used exogenous growth factors on embryo development up to the morula or blastocyst stage on Day 7, or blastocyst stage on Day 8. Moreover, medium supplemented with PDGF had fewer blastocysts than the control (p < or = 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Simultaneous in vitro fertilization and gamete intrafallopian transfer (GIFT).

During a 6-month period, a combination of gamete intrafallopian transfer (GIFT) and in vitro fertilization (IVF) was offered to all couples beginning an IVF treatment cycle in whom the wife had anatomically normal fallopian tubes. It was recommended to these couples that sufficient oocytes be reserved for insemination in vitro to determine whether the husband's spermatozoa could fertilize the wife's oocytes. During this interval, 16 couples underwent the combined IVF-GIFT procedure. All of the IVF-GIFT couples had at least two oocytes inseminated in vitro and at least two oocytes for GIFT. Of the 16 IVF-GIFT couples, only 1 (6.25%) achieved a clinical pregnancy. More important, 50% (8/16) of the IVF-GIFT couples had no oocytes fertilized in vitro. With the information concerning lack of fertilization in vitro, appropriate recommendations concerning future fertility management can be made. If the same couples had undergone the GIFT procedure alone, without additional oocytes fertilized in vitro, this information would not have been obtained.

Fallopian Tubes↗
PubMed2005

Successful pig embryonic development in vitro outside a CO2 gas-regulated incubator: effects of pH and osmolality.

We investigated the effects of HEPES in the medium (to maintain pH) and paraffin oil covering the medium (to maintain osmolality) on the developmental ability of porcine embryos produced in vitro using tightly closed glass tubes in the absence of a CO2 gas-regulated incubator. Putative porcine zygotes obtained by in vitro fertilization (IVF) of in vitro-matured (IVM) oocytes (day of IVF=Day 0) were cultured in 5% CO2 gas-equilibrated NCSU-37 media containing pyruvate and lactate during Days 0-2, and glucose during Days 2-6, in open glass tubes in a CO2 incubator or tightly closed glass tubes without a CO2 incubator at 38.5 degrees C. The following four media were used: (1) medium covered with paraffin oil and supplemented with HEPES; (2) medium covered with paraffin oil but with no HEPES supplementation; (3) medium not covered with paraffin oil but supplemented with HEPES; (4) medium not covered with paraffin oil and with no HEPES supplementation. As a control group, zygotes were cultured in medium with neither paraffin oil coverage nor HEPES supplementation using a four-well dish in a CO2 gas-regulated incubator. After culture, the osmolality in each of the four closed conditions was maintained at approximately 285-286 mOsm, lower (P<0.05) than that in the control (291 mOsm). In the two HEPES-supplemented media groups in the closed-tube system, the pH was maintained at 7.5-7.7, and the blastocyst development rates (15.5% in non-oil covered and 18.5% in oil covered group) did not differ significantly from that of the control (20.2%), although the mean cell numbers in the blastocysts in the two closed-tube condition groups (28.2 and 33.0) were lower (P<0.05) than in the control (43.5). In contrast, the pH was higher in the two groups without HEPES supplementation (approximately 8.0) than the control (7.4; P<0.05), and the blastocyst development rates (10.9% in non-oil covered and 7.5% in oil covered group) or total cell numbers in the blastocyst (24.8 and 28.7) in the two non-HEPES groups were drastically decreased (P<0.05) compared to those in the control (20.2% and 43.5). These results suggested that maintenance of pH is important for successful in vitro porcine embryo culture under closed-air conditions, whereas the range of osmolality that suits embryo development is not limited to a small range. Furthermore, blastocyst production was possible in a glass tube without a CO2 incubator, although blastocyst quality was lower compared to those produced in an incubator.

Animals↗
PubMed1991

Early pregnancy wastage in IVF (in vitro fertilization) patients.

Human in vitro fertilization is characterized by a low efficiency of implantation. Possible mechanisms for pregnancy loss are discussed. Embryo viability or quality, abnormal implantation, and delayed or absent corpus luteum rescue may all play a role in pregnancy wastage. Defining the possible mechanism for these losses may allow hormonal treatment to correct specific abnormalities.

Abortion, Spontaneous↗
PubMed1991

Preincubation of human granulosa cells with gonadotrophin prevents the cloprostenol-induced inhibition of progesterone production.

Human granulosa cells, from women undergoing ovum collection for in-vitro fertilization (IVF), will luteinize in vitro and provide a model for investigating the antigonadotrophic action of a prostaglandin F2 alpha (PGF2 alpha) analogue, cloprostenol, on granulosa-derived luteal cells. The granulosa cells were cultured in a defined medium and exposed to treatments during a preincubation period of 0 to 3 days and a final incubation with low density lipoprotein (LDL) from days 3 to 4. In the absence of human chorionic gonadotrophin (HCG), progesterone production was low, whereas exposure to HCG in the final incubation resulted in a 10-fold increase in progesterone concentrations. The inclusion of cloprostenol with HCG in the final incubation significantly (P less than 0.05) inhibited HCG-stimulated progesterone production. Exposure to HCG during the preincubation prevented the antigonadotrophic action of cloprostenol in the final incubation. The antigonadotrophic action of cloprostenol was retained when the granulosa cells were exposed to cloprostenol during the preincubation. Omission of LDL from the final incubation lowered the production of progesterone but the pattern of responses to HCG and cloprostenol were similar. Prevention of the antigonadotrophic action of cloprostenol after exposure to HCG may be a mechanism through which chorionic gonadotrophin can prevent regression of the corpus luteum in early pregnancy. Cloprostenol does not appear to inhibit LDL-stimulated steroidogenesis in human granulosa cells.

Cells, Cultured↗
PubMed2003

Energy metabolism in pig oocytes and early embryos.

Pig oocytes and embryos differ from those of other species in having a large quantity of endogenous lipid, a potential role for which has yet to be identified. In the present study, the hypothesis that endogenous triglyceride acts as a metabolic substrate during in vitro maturation and early embryo development was tested. Embryos were produced by in vitro fertilization (IVF) of in vitro-matured, abattoir-derived immature oocytes, cultured in medium NCSU23 up to the blastocyst stage. The triglyceride content of single oocytes and embryos was measured throughout development. Oxygen and glucose consumption and the formation of lactate were measured non-invasively over the same period, enabling total ATP production to be calculated. The triglyceride content of oocytes before maturation (135+/-4.9 ng) decreased by 13 ng (P<0.05) during in vitro maturation, but there was no apparent change in triglyceride content during embryo development (117.68 ng). Oxygen consumption was low throughout embryo cleavage before reaching a peak at the blastocyst stage (P<0.01), a pattern similar to that seen in other mammals studied. Glucose consumption and lactate production were also at a maximum at the blastocyst stage (P<0.05). These data indicate that pig oocytes may use endogenous triglyceride as an energy source during in vitro maturation and that most (91-97%) of the ATP produced during embryo development comes from oxidative phosphorylation. The high exogenous glucose concentration in NCSU23 (5.5 mmol l(-1)) may be needed to form pyruvate, which in turn, produces oxaloacetate, which is required to prime the tricarboxylic acid cycle. However, the reason for the high lipid content in early pig embryos remains to be elucidated.

Adenosine Triphosphate↗
PubMed2001

In vitro fertilization causes epigenetic modifications to the onset of gene expression from the zygotic genome in mice.

The effect of in vitro fertilization (IVF) and culture of mouse preimplantation embryos in vitro on the onset of expression of insulin-like growth factor 1 (IGF-1) ligand and receptor, insulin ligand and receptor, alpha-transforming growth factor (alpha-TGF) ligand, PAF:acetylhydrolase 1b (Pafah1b; alpha(1), alpha(2), and beta subunits of the enzyme), and the transcription requiring complex proteins (TRC) was examined. The IGF-1 ligand was detected in preimplantation embryos by immunofluorescence at all developmental stages tested. However, IVF and culture significantly reduced the amount of protein detected in the 8-cell embryo and blastocyst (P: < 0.001), and this was due to a delayed onset of expression of the mRNA for IGF-1 ligand from the zygotic genome. The expression of the alpha(1) subunit of Pafah1b was first detected at the 2-cell stage in fresh embryos, but expression was significantly retarded (P: < 0.001) when IVF and ISF (in situ-fertilized) zygotes were cultured in vitro. In vitro fertilization or ISF did not delay the onset of expression of TRC nor mRNA for the IGF-1 receptor, insulin receptor, alpha(2) or beta subunit of Pafah1b, nor did they effect alpha-TGF protein synthesis. Thus, IVF causes epigenetic modification in the normal pattern of expression of some but not all genes involved in normal embryo growth and survival.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗