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PubMed1996

Polyploidy and failed fertilization in in-vitro fertilization are related to patient's age and gamete quality.

A review of 392 cycles of in-vitro fertilization (IVF) was carried out in order to identify whether any factors such as sperm concentration at insemination, sperm motility or morphology, oocyte grading, number of oocytes retrieved, patient age, follicular stimulation protocols or duration of follicular growth, could be associated with the incidence of polyploidy or completely failed fertilization. The majority of polypronuclear fertilizations occurred in mature oocytes and in patients < 37 years of age and the incidence of polyploidy was strongly associated with fertilization and pregnancy rates. Fertilization rates were highest with mature oocytes. A significant linear trend was observed with failed fertilization and sperm concentration, morphology and motility. High rates of failed fertilization were found in patients > 37 years of age and with one to five oocytes retrieved. No significant difference was seen in stimulation protocols and oocyte grading between oocytes that fertilized and those that did not. Cycles with good sperm morphology and motility, the presence of mature oocytes, the retrieval of a large number of oocytes and younger maternal age seem to provide the best chance for IVF success.

Adult↗
PubMedDate not supplied

Hemizona index (HZI) demonstrates excellent predictability when evaluating sperm fertilizing capacity in in vitro fertilization patients.

The objective of this work was to assess the power of the hemizona assay to predict in vitro fertilization (IVF) results using receiving operating characteristics (ROC) analysis. In a tertiary medical center setting, 94 couples undergoing IVF were included in the study comparing their fertilization results to sperm function during the hemizona assay. Using ROC analysis, fertilization in IVF (mature oocytes only) was compared to performance of the sperm in the sperm-zona pellucida binding test as expressed by hemizona index (HZI; [number of test sperm bound/number of control sperm bound] x 100). The IVF results were as follows: 32 patients had no fertilization (0%), 55 patients fertilized all of the oocytes (100%), and 7 patients fertilized between 14% and 85% of the oocytes. Modified ROC analysis was used to determine the best threshold for fertilization success or failure, depending upon how the data are viewed. A HZI of 40-43 separated the 100% fertilization group from the other groups, with a true positive rate of 87% and a true negative of 83%. Conversely, a HZI of 4-5 separated the 0% fertilization versus partial or total fertilization, with a true positive rate of 100% and true negative rate of 71%. Sperm performance in the hemizona assay as measured by the HZI reflects performance in IVF and benefits couple evaluation and counseling.

Female↗
PubMed1995

Two embryonal cancers after in vitro fertilization.

BACKGROUND: In vitro fertilization is not considered to be associated with an increased rate of pediatric malignancies, and only three have been reported in the literature. Two additional rare pediatric tumors in children conceived through this technique are reported. METHODS: Two children 12 and 18 months of age, developed hepatoblastoma and clear cell sarcoma of the kidney, respectively. They were both products of uneventful pregnancies induced by in vitro fertilization. No other environmental, prenatal, or family factor was found. RESULTS: The first child died after a failed remission induction with cisplatin, doxorubicin, and vincristine, whereas the second child is alive with no evidence of disease 18 months after diagnosis and treatment according to NWTS protocol. CONCLUSIONS: A possible association between in vitro fertilization and pediatric malignancies is suggested.

Antibiotics, Antineoplastic↗
PubMed2004

Cryopreserved sibling oocytes and intracytoplasmic sperm injection rescue unexpectedly poor fertilization in conventional in vitro fertilization.

PURPOSE: To report a successful pregnancy from cryopreserved sibling oocytes and intracytoplasmic sperm injection (ICSI) for an infertile couple with an unexpectedly low fertilization rate in the fresh in vitro fertilization (IVF) cycle. METHODS: The woman had bilateral tubal obstruction and polycystic ovarian syndrome. The man had normal semen parameters. The couple underwent a cycle of controlled ovarian hyperstimulation in that 20 oocytes were retrieved. Twelve oocytes were conventionally inseminated and eight were cryopreserved using a slow freezing method. However, only one oocyte was fertilized, and no pregnancy was achieved. In the next cycle, the frozen oocytes were thawed and ICSI was performed. RESULTS: After thawing, seven oocytes (88%) survived and one was damaged. Six were at the metaphase II stage and were injected. Five (83%) achieved normal fertilization, and all of them cleaved (100%). After replacement of the embryos, a singleton pregnancy developed. A healthy female baby was delivered at term. Karyotyping revealed 46, XX. CONCLUSIONS: In addition to well-known indications, cryopreservation of excess sibling oocytes for patients receiving IVF has a possible advantage of preventing unexpectedly low fertilization rate or fertilization failures.

Adult↗
PubMed1999

[Results of in vitro fertilization therapy methods in Germany in 1996--the German In Vitro Fertilization Registry].

We believe that on the IVF subject, which is nowadays applied exept a standard method in the world. In the next years we could expect further improvement in our success++. It is also to anticipate that next changes in the development of assistance-reproduction will be: 1. Freezing of ovums, 2. To get in vitro-meture of the immature cells, 3. Transplantation of ovarium tissue, 4. Transfer of the blastocyst, 5. Transfer of cytoplasma or the cell nucleus. We recommend that the IVF/ICSI--therapy and birth of children has to be registrated in one central register.

Adult↗
PubMed1992

Effects of superoxide dismutase on mouse in vitro fertilization and embryo culture system.

PURPOSE: We recently found that, for mouse embryos fertilized in vivo, the two-cell block could be attenuated by adding superoxide dismutase (SOD), a scavenger of superoxide radicals, to the culture medium. In this study, we evaluated the effects of SOD on the process of fertilization and on the further development of the embryos fertilized in vitro. METHODS: We performed incubation of mouse epididymal spermatozoa, in vitro fertilization, and further cultivation in Biggers-Whitten-Whittingham's medium supplemented with various concentrations of Cu.Zn-SOD. RESULTS: High concentrations (2000 micrograms/ml or more) of SOD prevented loss of motility in mouse sperm over time. The addition of SOD (less than 2000 micrograms/ml) to the basic medium showed no significant difference in the fertilization rate. Also, no significant difference was observed in the rate of polyspermy or parthenogenesis between the basic and the SOD-supplemented media. However, 18% of the two-cell-stage embryos developed to the expanded blastocyst stage in the 500 micrograms/ml SOD-supplemented medium, while no blastocysts were found in the basic medium. Furthermore, the addition of SOD 7 hr after insemination increased the expanded blastocyst rate (28%). CONCLUSIONS: These results indicate that the addition of SOD exerts a protecting effect from oxidative stress both on sperm viability and on the development of embryos fertilized in vitro as well as in vivo, while its addition showed no effect on the process of fertilization.

Animals↗
PubMed1991

Sex and development in bovine in-vitro fertilized embryos.

In vitro fertilized bovine embryos were karyotyped at Day 7 and Day 8 post insemination. The results showed that the percentage of males (sex ratio) was dependent on the developmental stage. Among expanded blastocysts (the most advanced embryos), the sex ratio was 100%, followed by 89% for expanding blastocysts, 75% for blastocysts, and 40% for young blastocysts, all analyzed at Day 7. For embryos karyotyped at Day 8, the sex ratio was 20%. The average mitotic index for all in vitro fertilized embryos was 3.5%. These results suggest that the apparent relationship between sex and developmental rate could be used as a method for noninvasive sexing of in vitro fertilized embryos.

Journal Article↗
PubMed1988

Decreased embryonic survival of in-vitro fertilized oocytes in rats is due to retardation of preimplantation development.

Immature female rats (60-65 g) were injected with 4 i.u. PMSG on Day -2 and allocated to 3 groups. On the evening of Day 0, rats in Groups I and II were allowed to mate. Embryos were collected on Day 4 (Group I, control morulae) or Day 5 (Group II, control blastocysts) and were transferred into the oviduct or uterine horn of Day-4 pregnant recipient rats. On the transfer side of the recipients, the bursa had been peeled from around the ovary to prevent endogenous oocytes from entering the oviduct. For Group III, unmated donors were killed 65-67 h after PMSG injection. Ovulated oocytes recovered from the oviducts were fertilized in vitro and transferred 16-18 h later. Embryos developing from in-vitro fertilized (IVF) oocytes were recovered on Day 5, separated into morulae (Group IIIm) and blastocysts (Group IIIb) and transferred into Day-4 pregnant recipients similar to control embryos. Some embryos from each group were used to determine the mean number of cells/embryo. Embryo recipients were killed on Day 20. After transfer, the development of IVF oocytes was retarded compared to control embryos. IVF morulae contained significantly fewer cells/embryo than did control morulae but were able to implant and grow to fetuses, in proportions similar to controls, if transferred into the oviduct of the recipients. These results suggest that the developmental potential of rat oocytes fertilized in vitro is limited due to asynchrony between the embryo and the uterine environment at the time of implantation, rather than possible defects incurred by the oocyte during the fertilization procedure.

Animals↗
PubMed1992

Investigation of means to improve rates of fertilization in in vitro matured/in vitro fertilized bovine oocytes.

Experiments were conducted with 5,979 oocytes to determine whether detaching some of the cumulus cells from oocytes either before or after maturation would improve the fertilization rate and proportion of oocytes that developed to expanded blastocysts. Oocytes were aspirated from ovaries of slaughtered cows and matured, fertilized and cultured in vitro. Pipetting immature oocytes before maturation to detach some of the cumulus, with all cumulus cells left in the maturation wells, significantly increased fertilization rates, especially of oocytes that initially had a full cumulus investment. In further experiments, pipetting oocytes either before or after maturation to detach most of the cumulus, or treating with hyaluronidase after maturation to disperse the cumulus, significantly increased fertilization rates and proportions of oocytes developing to expanded blastocysts.

Journal Article↗
PubMed1983

Nonhuman primate in vitro fertilization: biochemical changes associated with embryonic development.

Little research has focused on the biochemistry of primate embryos fertilized in vitro. These studies were designed to examine alterations in cellular incorporation of leucine, uptake of steroid hormones, and overall viability of such embryos. The results may be applied to certain investigations of embryonic development in humans. Squirrel monkey ova and embryos were incubated in medium supplemented with either 3H-leucine, 3H-17 beta-estradiol (E2), or 3H-progesterone (P) and processed for autoradiographic analysis or assayed for radioactivity. Cellular incorporation of 3H-leucine (as an indication of protein synthesis), declined with oocyte maturation in vitro and remained constant at in vitro fertilization (IVF). The use of the vital dyes trypan blue and fluorescein diacetate showed high correlations with the cellular incorporation of 3H-leucine by ova, therefore demonstrating viability. Uptake of E2 and P increased at IVF. The results indicate normal metabolic development of primate embryos fertilized in vitro that is comparable to preimplantation development of other mammalian species.

Animals↗
PubMed1987

The in vitro fertilization of supernumerary oocytes in a gamete intrafallopian transfer program.

Gamete intrafallopian transfer (GIFT) was performed on a series of 42 couples in whom four oocytes were replaced into the fallopian tubes and the remaining oocytes were inseminated with 100,000 motile spermatozoa. Fertilization in vitro was assessed 16 to 20 hours later. An overall pregnancy rate after GIFT of 40.0% (12/30) in normospermic couples and 16.7% (2/12) in oligospermic couples was seen. Supernumerary oocytes were submitted to in vitro fertilization (IVF) and the findings related to the likelihood of pregnancy. No correlation between IVF and the likelihood of GIFT pregnancies was found in either the oligospermic or normospermic couples. A similar proportion of pregnancies was seen in couples with or without evidence of fertilization of the supernumerary oocytes in both the oligospermic and normospermic couples. However, a high proportion of normospermic couples with no evidence of fertilization had only one oocyte remaining for insemination. These results suggest that the failure of low numbers of supernumerary oocytes to fertilize in vitro after GIFT relates to oocyte selection criteria and does not mean a reduced chance of conception in that treatment cycle.

Fallopian Tubes↗
PubMed1988

Ultrastructural abnormalities of in vitro fertilization of in vitro matured bovine oocytes.

Cumulus-oocyte complexes collected from cows at an abattoir by aspiration from small (1-6 mm) antral follicles were matured and inseminated in vitro. At different time intervals after insemination the ova were processed for transmission electron microscopy. Up to and including 6 h after insemination all ova were unfertilized, and their cortical granules were more or less clustered. At 6 h acrosome reaction of spermatozoa was observed on the surface of the zona pellucida. At 8 h the first fertilized ovum appeared and the first fully developed spherical pronucleus was observed, at 20 h the first apposition of pronuclei was seen, and at 40 h divisions were ongoing or completed. More than one third of the fertilized ova showed polyspermic penetration of the zona pellucida, and in most of these ova different developmental stages of supernumerary pronucleus formation were observed in the ooplasm. Abnormal cortical granule release was seen in approximately half of the fertilized ova, and it was more frequent in ova with polyspermic as opposed to monospermic penetration of the zona pellucida.

Animals↗
PubMed1998

Creatine kinase activity in human spermatozoa and seminal plasma lacks predictive value for male fertility in in vitro fertilization.

OBJECTIVE: To examine the predictive value of creatine kinase in human spermatozoa and seminal plasma from patients treated in an IVF program. DESIGN: Prospective, blind clinical study. SETTING: Male infertility clinic. PATIENT(S): Seventy-three patients and 32 fathers (age, <45 years). INTERVENTION(S): Determination of creatine kinase activity in seminal plasma, washed spermatozoa, and swim-up purified spermatozoa from patients treated in an IVF program. MAIN OUTCOME MEASURE(S): Creatine kinase activity in seminal plasma, washed spermatozoa, and swim-up purified spermatozoa. RESULT(S): Creatine kinase activity in washed spermatozoa correlated significantly with normal sperm morphology. No significant correlations were found between creatine kinase activity or creatine kinase isoenzyme ratio in seminal plasma, washed spermatozoa, or swim-up purified spermatozoa and success in the IVF program. The percentage of spermatozoa with normal morphology was significantly lower in patients whose sperm did not fertilize than in patients whose sperm did fertilize oocytes and in the control group of proven fathers. CONCLUSION(S): Total creatine kinase activity and creatine kinase isoenzyme distribution are not sperm function markers for prediction of male fertility in IVF treatment.

Adult↗
PubMed1986

Superovulation of mice with human menopausal gonadotropin or pure follicle-stimulating hormone in combination with human chorionic gonadotropin and the effects of oocyte aging on in vitro fertilization.

The response of female mice of F1 hybrids (CBA x C57/BL) to superovulatory doses of human menopausal gonadotropin (hMG) or pure follicle-stimulating hormone (FSH) in combination with human chorionic gonadotropin (hCG) was studied. Furthermore, the effect of oocyte aging in vivo on the subsequent rate of fertilization in vitro was also investigated. The oocytes were collected at 12, 18, and 24 hr after hCG injection and in vitro fertilization (IVF) was carried out in T6 medium. A higher proportion of animals responded to hMG stimulation (32/70) compared to pure FSH (15/66). Furthermore, hMG gave a higher oocyte recovery (454/32) than pure FSH (77/15). Fertilization rates of 57.8, 51.5, and 53.5% were obtained for the 12-, 18-, and 24-hr groups, respectively, after correction for parthenogenetic division of oocytes in the controls. No significant differences in fertilization rates were observed among the three time intervals used in recovering oocytes. However, as the degeneration and parthenogenetic division increased with the delay in collection of oocytes, 12 hr post-hCG injection was the best time to collect oocytes to obtain optimum results in in vitro fertilization.

Animals↗
PubMed1976

In vitro fertilization, culture, and transfer of rabbit ova.

Ovulated rabbit oocytes were fertilized in vitro in chemically defined media supplemented with bovine serum albumin and either cultured up to the expanding blastocyst stage or transferred to recipients after varying periods of culture. Embryos transferred after up to 72 hours of in vitro culture were born as viable young. Oocytes from young virgin does were superior to oocytes from nonvirgin does for the purpose of in vitro fertilization (54% versus 26% fertilized, P less than 0.01). Capacitated sperm from artificially inseminated capacitators resulted in fertilization rates slightly lower than those from naturally mated does (46% versus 57% fertilized, P less than 0.025). Removal of cumulus and corona cells from oocytes with hyaluronidase and repeated aspiration through a fine pipette resulted in lowered fertilization rates (51% versus 73%, P less than 0.025). Linbro Disposo Tray wells were as good as glass tissue-culture dishes for the in vitro mixing of gametes and were more convenient to use. Modified Ham's F10 medium was used to culture the in vitro-fertilized embryos. However, when a modified Brackett's medium was used instead of modified Ham's F10 for the initial 4-hour period after mixing gametes, more oocytes were fertilized (52% versus 28%, P less than 0.01).

Animals↗
PubMed1995

Cigarette smoking at time of in vitro fertilization cycle initiation has negative effect on in vitro fertilization-embryo transfer success rate.

PURPOSE: To assess cigarette smoking of female patients prior to starting in vitro fertilization cycle and possible affect on subsequent in vitro fertilization-embryo transfer outcome. METHODS: Retrospective study involving 340 consecutive patient questionnaires filled out at time of in vitro fertilization program entry. Only cycles resulting in embryo transfer after transvaginal ultrasound directed ovum retrieval (n = 253) were considered. The three patient pregnancy outcomes of not pregnant, spontaneous abortion, and live birth, were cross-referenced with smoking and nonsmoking patients. RESULTS: No significant difference found in overall pregnancy rate per embryo transfer for smokers (35%) vs nonsmokers (31%). However, the abortion rate was significantly higher for the smokers (73%) vs nonsmokers (24%) with a P value < 0.001. CONCLUSIONS: Results suggest preentry in vitro fertilization cycle cigarette smoking has adverse affect on potential pregnancy outcome by increasing spontaneous abortion. Preconception health consultation concerning adverse cigarette smoking effects should be implemented prior to program entry.

Abortion, Spontaneous↗