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Human Fallopian tube epithelial cell co-culture increases fertilization rates in male factor infertility but not in tubal or unexplained infertility.

In order to investigate the effect of human Fallopian tube epithelial cell co-culture on fertilization and cleavage rates in tubal, male and unexplained infertility, oocytes collected from 91 patients were randomized to wells containing Fallopian tube epithelial cell monolayers or conventional culture medium, and inseminated with spermatozoa. Fertilization and cleavage were assessed at 18 and 52 h, respectively. Co-culture significantly increased the fertilization rates over the control values in male infertility (41.67 versus 23.43%, P = 0.00005), but not in tubal infertility (69.33 versus 67.93%) or unexplained infertility (65.93 versus 54.36%). Cleavage rates were not different in co-culture and conventional in-vitro fertilization systems in any of the infertility subgroups. The number of blastomeres was significantly higher in the co-culture group on the day of embryo transfer (3.63 +/- 1.12 versus 3.04 +/- 1.26, P < 0.001). Pregnancy rates were similar in all infertility subgroups. There was no significant association between the number of co-cultured embryos transferred and the pregnancy, abortion and multiple pregnancy rates. It was concluded that human Fallopian tube epithelial cell co-culture clearly improves fertilization rates in male infertility but not in tubal or unexplained infertility. Improved fertilization rates in co-culture may be due to positive effect of co-culture on impaired sperm function.

Blastomeres↗

Effects of activation timing on the fertilization rate and early embryo development in porcine ROSI procedure.

PURPOSE: This study was undertaken to evaluate the optimal exposure time for nuclear envelope breakdown (NEBD) after injection of a round spermatid and to investigate the effect of oocyte activation timing on the fertilization rate and early embryo development in porcine round spermatid injection procedure. METHODS: Injected oocytes were fixed at 0.5, 1, 2, 3, and 4 h before activation, and NEBD state was examined. The three groups of oocytes were activated before and after the injection of spermatid using a single direct current pulse (100 V/mm, 50 micros): group 1) at 2 h before the injection (pre), group 2) within 0.5 h after the injection (immediate), and group 3) at 2 h after the injection (post). Activated oocytes were cultured and pronucleus formation and blastocyst development was evaluated at 15-18 h and 7-8 days after the injection, respectively. RESULTS: The proportion of oocytes with NEBD significantly increased in the groups with over 2 h of exposure time (p < 0.05) and oocyte with premature chromosome condensation began to appear 3 h after the injection. Normal fertilization and development rate to the blastocyst stage were significantly higher in the post group than in those of the pre or immediate group (p < 0.05). CONCLUSION: The optimal exposure time for NEBD is 2 h after the injection, and activation in 2 h after round spermatid injection improved the normal fertilization and early embryo development rate.

Animals↗

Fertilization rate in couples with unexplained infertility.

A group of 24 couples with unexplained infertility was scheduled for in-vitro fertilization and tubal embryo transfer between May 1989 and September 1990. In the same period, in-vitro fertilization and intrauterine transfer of embryos was planned in a control group of 44 women with tubal infertility. The mean age and duration of infertility were similar in both groups and the same scheme of ovarian stimulation was used. No statistically significant difference was obtained comparing oestradiol levels and numbers of mature oocytes retrieved between the group of patients with unexplained infertility and those with tubal infertility. The fertilization rate of the oocytes obtained from women with unexplained infertility (60.4%) was significantly lower (P less than 0.001) than that of the oocytes obtained from patients with tubal infertility (87.3%). There was no statistically significant difference in the cleavage rates between patients with unexplained infertility and those with tubal infertility. It is concluded that lack of fertilization is an unexplored cause of infertility in couples with unexplained infertility.

Adult↗

Effect of level of endophyte infection, nitrogen fertilization rate, grazing period, and paddock exchange on some chemical properties of four bovine tissues.

Three grazing experiments were conducted to determine the effect of level of endophyte infection, rate of N fertilization of tall fescue grass, grazing period, and paddock exchange on selected chemical properties of four bovine carcass tissues. Samples of semitendinosus and longissimus muscle and of subcutaneous and perinephric adipose tissue were excised from the left side of each carcass. In Exp. 1, percentage of stearic acid was higher (P less than .05) and percentages of palmitoleic and oleic acid were lower (P less than .05) in all tissues from steers grazed on LELN Au-Triumph fescue than in tissues from steers grazed on LELN KY-31 fescue. Grazing periods of 175 or 245 d or paddock exchange (Exp. 2) had no significant effect on proximate composition of semitendinosus and longissimus muscles; however, moisture content was higher (P less than .05) in the semitendinosus muscle from steers grazed on 100% (100E) endophyte-infected KY-31 fescue. Forage treatment, grazing period, and paddock exchange (Exp. 2 and 3) had no significant effect on percentage of saturated fatty acids in the subcutaneous, semitendinosus, and longissimus tissue samples. In Exp. 2 and 3, percentages of saturated fatty acids were higher (P less than .05) in subcutaneous and perinephric adipose tissue samples from steers grazed on 100E than in samples from steers grazed on zero endophyte (OE) fescue.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

Chromosome analysis in human oocytes remaining unfertilized after in-vitro insemination: effect of maternal age and fertilization rate.

The incidence of chromosomal abnormalities was studied in 719 unfertilized human oocytes obtained from our in-vitro fertilization (IVF) programme. To make chromosome preparations, a gradual fixation/air-drying method was utilized. Of 388 oocytes successfully karyotyped, 70 (18.0%) were abnormal. The abnormalities included 33 aneuploidies (8.5%) (14 hyperhaploidies and 19 hypohaploidies), 25 diploidies (6.4%) and 15 structural abnormalities (3.9%), three of them being accompanied by aneuploidy. Of the 33 aneuploidies, 16 (48.5%) showed the loss or gain of dyads (so-called non-disjunction), while 17 (51.5%) showed the loss or gain of monads (so-called predivision). There was no maternal age-dependent increase in the incidence of aneuploidy. Unfertilized oocytes from patients with a high fertilization rate (>25%) had a significantly higher (11.4%, P < 0.05) incidence of diploidy compared with the oocytes from the remaining patients (4.3 and 4.0%), suggesting that diploid oocytes might have a lower fertilizing ability.

Adult↗

Total immunoreactive alpha-inhibin in human seminal plasma, sperm quality, and in vitro fertilization rates.

OBJECTIVE: To assess the relationship between the concentrations of total immunoreactive alpha-inhibin in human seminal plasma, sperm quality, and in vitro fertilization (IVF) rates. SETTING: Medical school hospital. DESIGN: Prospective study. PATIENTS: 72 consecutive couples undergoing IVF with husband's sperm in which at least 4 oocytes were obtained. MAIN OUTCOME MEASURES: Alpha-inhibin determination by means of enzyme-linked immunosorbent assay, sperm analysis for density and motility, IVF rates. RESULTS: Total immunoreactive alpha-inhibin levels were not correlated with IVF rates. Nor were there any differences in fertilization rates in the different groups. Lower inhibin levels were found in oligozoospermic cases. Lower sperm counts were found in cases with inhibin values below the median value, and also with respect to various other cutoffs. No correlation was found between inhibin levels and any of the sperm variables measured. CONCLUSION: Total immunoreactive alpha-inhibin is not a prognostic factor in IVF cycles. However, an association was found between inhibin and sperm concentration, suggesting that alpha-inhibin in the human seminal fluid could be a marker for some aspects of spermatogenesis.

Adult↗

[Concentrations of MMP-1, TIMP-1, MMP-1/TIMP-1 and I CTP complexes in follicular fluid as related to fertilization rate in women treated with in-vitro fertilization].

OBJECTIVES: Metalloproteinases and their inhibitors appear to control connective tissue remodelling during follicular rupture. The aim of the study was to establish if oocytes fertilisation rate after ovulation induction depends on the concentrations of MMP-1, its inhibitor TIMP-1, MMP-1/TIMP-1 complex and I CTP in follicular fluid (FF). MATERIAL AND METHODS: FF were collected from 37 infertile patients undergoing ovulation induction using either short or long protocol. FF was obtained 36 hours after administration of hCG (Pregnyl). The level of MMP-1, TIMP-1, MMP-1/TIMP-1 complex were measured using ELISA kits and I CTP, E2, FSH, LH, using RIA assay kits. RESULTS: Statistically significant (p < 0.05) difference was found in TIMP-1, E2, and FSH concentration, being higher in the group with more than 75% fertilisation rate: TIMP-1 728.8 + 100.1 ng/ml vs 666.3 + 94.5 ng/ml; E2 477.3 +/- 160.0 ng/ml vs 368.0 +/- 190.0 ng/ml and FSH 7.27 +/- 1.45 mIU/ml vs 6.24 +/- 1.6 mIU/ml. CONCLUSIONS: Statistically significant increase in TIMP-1 concentration observed among patients with fertilisation rate above 75% indicates an important role of this substance in ovulation process.

Adult↗

Effects of immunization against bone morphogenetic protein 15 and growth differentiation factor 9 on ovulation rate, fertilization, and pregnancy in ewes.

Immunization of ewes against growth differentiation factor 9 (GDF9) or bone morphogenetic protein 15 (BMP15) can lead to an increased ovulation rate; however, it is not known whether normal pregnancies occur following such treatments. The aims of the present study were to determine the effects of a short-term immunization regimen against BMP15 and GDF9 on ovulation rate, fertilization of released oocytes, the ability of fertilized oocytes to undergo normal fetal development, and the ability of immunized ewes to carry a pregnancy to term. Ewes were given a primary and booster immunization against keyhole limpet hemocyanin (KLH; control, n = 50), a GDF9-specific peptide conjugated to KLH (GDF9, n = 30), or a BMP15-specific peptide conjugated to KLH (BMP15, n = 30). The estrous cycles of all ewes were synchronized, and ewes were joined with fertile rams approximately 14 days after the booster immunization. The number of corpora lutea was determined by laparoscopy 3-4 days following mating. Subsequently, about one-half of the ewes in each group underwent an embryo transfer procedure 4-6 days following mating, with the embryos being transferred to synchronized, nonimmunized recipients. The remaining ewes were allowed to carry their pregnancies to term. Short-term immunization against either BMP15 or GDF9 peptides resulted in an increase in ovulation rate with no apparent detrimental affects on fertilization of released oocytes, the ability of fertilized oocytes to undergo normal fetal development, or the ability of the immunized ewes to carry a pregnancy to term. Therefore, regulation of BMP15, GDF9, or both is potentially a new technique to enhance fecundity in some mammals.

Amino Acid Sequence↗

Preliminary counseling with age-related fertility rates does not influence whether women undergo reversal of sterilization.

Our objective was to assess the influence of preliminary counseling that provided information about age-related fertility rates on the decision of women to undergo reversal of sterilization. There was no apparent influence; this raises the question as to whether couples seeking fertility therapy base their decision making on factual information provided by the clinician.

Adult↗

Cryoprotection of human oocytes: inappropriate exposure to DMSO reduces fertilization rates.

Human oocytes were exposed to the cryoprotectant dimethyl-sulphoxide (DMSO) at either 4 or 37 degrees C. Subsequent fertilization of these oocytes showed that exposure to DMSO at 37 degrees C was associated with a greatly reduced fertilization rate when compared to untreated control oocytes, whereas no such reduction was seen in oocytes exposed to DMSO at 4 degrees C. The significance of these results for the potential cryopreservation of human oocytes is discussed.

Cryopreservation↗

Cystic fibrosis mutations impair the fertilization rate of epididymal sperm from men with congenital absence of the vas deferens.

One of the limiting factors for the successful treatment of male sterility due to congenital absence of the vas deferens (CAVD) is the low (< 20%) and extremely unpredictable rate of in-vitro fertilization of their epididymal spermatozoa. The recent demonstration that CAVD is a mild form of cystic fibrosis (CF) disease, almost exclusively involving the genital tract, has prompted us to investigate the hypothesis of whether there could be an association between particular CF genotype mutations and the in-vitro performance of epididymal sperm. In this study 63 patients with surgically confirmed diagnosis of CAVD undergoing microsurgical epididymal sperm aspiration (MESA) and in-vitro fertilization (IVF) were evaluated. The genetic screening was carried out on DNA extracted from peripheral lymphocytes and amplified by the polymerase chain reaction. A total of 12 mutations in the cystic fibrosis transmembrane regulator gene (CFTR), representing approximately 90% of the total known CF mutations, were tested. The presence or absence of mutations, as well as the type of mutation found, was correlated with the IVF and pregnancy rates. Of the 63 patients examined, 40 (64%) were positive to the CF screening and 23 were negative. In three cases no spermatozoa were found because of rete testis blockage. The difference in fertilization rates between patients testing positive and negative was highly significant (P = 0.000003), 13 versus 23%, respectively. In order to pinpoint which mutation could account for the most deleterious effect on the IVF results, we analysed these by keeping the patients separated by CF mutations.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Hormonal parameters in follicular fluid and the fertilization rate of in vitro cultured oocytes.

Oocytes and matched samples of follicular fluid were obtained from 52 preovulatory follicles aspirated laparoscopically for in vitro fertilization (IVF). Follicular fluid concentrations of estradiol (E2), progesterone (P), testosterone (T), prolactin (HPRL), prostaglandin F2 alpha (PGF2 alpha), prostaglandin E (PGE), protein content, and collagenolytic activity were measured and related to the fertilization rate of oocytes cultured in vitro. High concentrations of P and low levels of T and HPRL were associated with mature, fertilizable oocytes. Levels of PGF2 alpha, PGE, and follicular fluid protein concentrations were similar in both groups. Mean collagenolytic activity was increased in the fertilized oocytes, although no significant difference could be observed. Our data demonstrate a close association between follicular fluid steroid and HPRL concentrations and successful fertilization of oocytes.

Dinoprost↗

Effects of insemination-ovulation interval on fertilization rates and embryo characteristics in dairy cattle.

The objective of this study was to examine effects of the interval between insemination and ovulation on fertilization and embryo characteristics (quality scored as good, fair, poor and degenerate; morphology; number of cell cycles and accessory sperm number) in dairy cattle. Time of ovulation was assessed by ultrasonography (every 4h). Cows were artificially inseminated once between 36h before ovulation and 12h after ovulation. In total 122 oocytes/embryos were recovered 7d after ovulation. Insemination-ovulation interval (12h-intervals) affected fertilization and the percentages of good embryos. Fertilization rates were higher when AI was performed between 36-24 and 24-12h before ovulation (85% and 82%) compared to AI after ovulation (56%). AI between 24 and 12h before ovulation resulted in higher percentages of good embryos (68%) compared to AI after ovulation (6%). Insemination-ovulation interval had no effect on number of accessory sperm cells and number of cell cycles when corrected for embryo quality. This study showed that the insemination-ovulation interval with a high probability of fertilization is quite long (from 36 to 12h before ovulation). However, the insemination-ovulation interval in which this fertilized oocyte has a high probability of developing into a good embryo is shorter (24-12h before ovulation).

Animals↗