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PubMed1975

In vitro fertilization of rat and mouse eggs by ejaculated sperm and the effect of energy sources on in vitro fertilization of rat eggs.

In vitro fertilization of rat and mouse eggs by ejaculated or epididymal spermatozoa in chemically defined media was studied. Penetration rates by ejaculated sperm was very low (0 to 8%) in the rat, but 11 to 41% of eggs were penetrated by ejaculated sperm in the mouse. The optimal concentration of sperm for in vitro fertilization appears to be similar whether ejaculated or epididymal sperm were used. The time of sperm penetration in the mouse eggs, however, was delayed for one-half to one hour when ejaculated sperm were used. The importance of sodium pyruvate, sodium lactate and glucose in the medium containing bovine serum albumin for in vitro fertilization of rat eggs was examined. When rat eggs in cumulus clot were exposed to epididymal sperm preincubated for five hours, the presence of sodium pyruvate, sodium lactate and glucose was found to play an important role. When exposed to non-incubated epididymal sperm sodium pyruvate could be omitted without much decline of the fertilization rate. When the denuded eggs were exposed to non-incubated sperm, penetration rates were very low (0 and 5%) in the absence of pyruvate. It appears that although lactate, pyruvate and glucose are all important for in vitro fertilization of rat eggs, pyruvate can be supplied by the follicular cells surrounding the eggs.

Animals↗
PubMed1999

Pleiotropic effects of a nuclear restorer-of-fertility locus on mitochondrial transcripts in male-fertile and S male-sterile maize.

Cytoplasmic male sterility (CMS) is encoded by the plant mitochondrial genome and can be reversed by nuclear restorer-of-fertility(Rf) alleles. In the CMS-S system of maize, reproductive failure and fertility restoration are gametophytic, occurring during the starch-filling stages of pollen development. Transcripts of the CMS-S-associated mitochondrial open reading frames (orf355 and orf77) are present from the early stages of microspore development through the aborted pollen stage. To investigate the molecular basis of fertility restoration, we compared mitochondrial-transcript accumulation in aborting CMS-S pollen and in CMS-S pollen restored to fertility by the Rf3 nuclear allele. In the presence of the Rf3 allele, novel, shorter transcripts of the orf355-orf77, cob and atp6 mitochondrial genes were created, and the relative abundance of larger transcripts was decreased for each of these loci. The altered transcript patterns cosegregated with male fertility conditioned by the Rf3 allele. The novel cob and atp6 transcripts were also observed in leaf-tissues of both normal and S-cytoplasm plants carrying the Rf3 allele. These observations support the hypothesis that the Rf3 allele encodes, or regulates, a modifier of mitochondrial transcript (Mmt) activity that affects both CMS and essential mitochondrial gene transcripts.

Alleles↗
PubMed2006

The effect of cancer treatment on female fertility and strategies for preserving fertility.

Aggressive chemotherapy and radiotherapy in young patients with cancer has greatly enhanced the life expectancy of these patients, but these treatments often cause infertility because of the massive destruction of the ovarian reserve resulting in premature ovarian failure (POF). This review focuses on the effect of cancer treatments on fertility and on the various surgical and assisted-reproduction innovations that are available to provide the patient with the option of future pregnancies. As the emerging discipline of fertility preservation is steadily attracting increasing interest, developments in the near future promise to be very exciting. However, in everyday routine work, better interdisciplinary cooperation between gynecological and pediatric oncologists, surgeons, immunologists and endocrinologists is necessary so that individualized options for fertility preservation can be offered in advance of surgical procedures or cancer treatments. GnRH analog treatment can preserve fertility in some patients, but not in all. At present, cryopreservation of ovarian tissue appears as a very promising method of providing the cancer patient with a realistic chance of preserving fertility-a prospect that is also extremely important to patients for psychological reasons.

Adult↗
PubMed2001

Intracytoplasmic sperm injection increased fertilization and good-quality embryo formation in patients with non-male factor indications for in vitro fertilization: a prospective randomized study.

OBJECTIVE: To compare the fertilization rate and formation of good-quality embryos with conventional IVF and ICSI in patients with non-male factor infertility. DESIGN: Prospective controlled study. SETTING: Infertility clinic. PATIENT(S): Thirty-five patients with non-male factor infertility. INTERVENTION(S): Retrieved sibling oocytes were randomly assigned to conventional IVF or ICSI. Of sibling oocytes assigned to ICSI, only metaphase II oocytes were injected with sperm. MAIN OUTCOME MEASURE(S): Fertilization rate and formation of good-quality embryos per retrieved oocyte. RESULT(S): Per retrieved oocyte, ICSI resulted in better fertilization rate compared with conventional IVF (71.3% [134 of 188] vs. 57.2% [107 of 187]). Per retrieved oocyte, ICSI also resulted in better formation of good-quality embryos at 48 hours after retrieval compared with conventional IVF (64.4% [121 of 188] vs. 47.1% [88 of 187]). CONCLUSION(S): In IVF patients with non-male factor infertility, subjecting some sibling oocytes to ICSI increased the fertilization rate and formation of good-quality embryos per retrieved oocyte. It also avoided the problem of total fertilization failure in almost all cases.

Adult↗
PubMed1990

Are factors that influence oocyte fertilization also predictive? An assessment of 148 cycles of in vitro fertilization without gonadotropin stimulation.

Fertilization characteristics of 152 consecutively obtained oocytes in an in vitro fertilization (IVF) program employing only natural and clomiphene citrate-induced cycles were retrospectively analyzed. Fertilization occurred significantly more often (1) in women with tubal infertility, (2) in spontaneous cycles, and (3) in cases of secondary infertility. Grade I sperm motility from the original semen sample and the duration of infertility were also significant influencing factors. A similar sperm correlate was not identified on samples after sperm migration. Preovulatory follicular fluid steroids, progesterone (P), estradiol (E2), E2:P ratio, and luteinizing hormone (LH), as well as baseline plasma LH and the magnitude of the LH surge did not correlate with fertilization. However, when the identified factors were used to predict fertilization (discriminant analysis), only 58.3% of oocytes were correctly classified. This data supports the concept of performing IVF as a test in its own right.

Adult↗
PubMed1992

Is fertility-problem stress different? The dynamics of stress in fertile and infertile couples.

OBJECTIVE: To compare the dynamics of fertility-problem stress experienced by wives and husbands in infertile couples with the dynamics of stress from other sources experienced by members of couples presumed to be fertile. DESIGN: Relationships of stress to four marriage factors and four aspects of life quality (subjective well-being) are examined within a causal modeling framework using data from structured interviews. SETTING: Face-to-face interviews were conducted in study participants' homes. PARTICIPANTS: Wives and husbands from 157 couples with primary infertility and from 82 presumed-fertile couples were studied. MAIN OUTCOME MEASURES: Final outcome measures were four multi-item scales assessing life quality with regard to the marriage, own self-efficacy, own health and appearance, and life as a whole. Intervening outcome scales measured four marriage factors: marital conflict, sexual self-esteem, sexual dissatisfaction, and frequency of intercourse. RESULTS AND CONCLUSIONS: Higher levels of stress, regardless of whether that stress was from attempting to solve a fertility problem or another problem, were related to reduced marital functioning and decreased life quality. For husbands, the strengths of the linkages did not depend on the source of the stress. For wives, however, the causal model suggested that fertility-problem stress had stronger negative impacts on sense of sexual identity and self-efficacy than did stress from other problems (P less than 0.05). Stress from any source had more impact on the lives of wives than of husbands, more impact on satisfaction with self and general well-being than on satisfaction with the marriage or health, and affected life quality mostly indirectly through its impacts on the marriage factors.

Adult↗
PubMed1994

Effect of follicular size on oocyte retrieval, fertilization, cleavage, and embryo quality in in vitro fertilization cycles: a 6-year data collection.

OBJECTIVE: To determine the effect of follicular size, including the size of the leading follicle, on oocyte retrieval, fertilization, cleavage, and embryo quality in IVF cycles based on a large data collection. DESIGN: Retrospective analysis of 1,109 IVF cycles between 1987 and 1993 at the Hospital of the University of Pennsylvania including 606 patients ranging in age from 23 to 49 years. RESULTS: Follicles with a volume < or = 1 mL show a significantly lower oocyte recovery rate than follicles with a volume of > 1 mL. The highest recovery rate (83.5%) was found in follicles with a volume of 3 to 4 mL. Above a follicular volume of 7 mL, the oocyte recovery drops below that observed for follicles between 1 and 7 mL. Fertilization and cleavage rates were also higher in oocytes obtained from follicles > 1 mL compared with follicles < or = 1 mL. Although fertilization rates were fairly stable above volumes of 1 mL, cleavage rates continued to rise to a peak percentage of 92% with volumes between 6 and 7 mL. Leading follicle size did not have an effect on fertilization and cleavage rates of cohort oocytes. Embryo quality was not influenced significantly by follicular volume. CONCLUSION: Based on this evaluation of a large number of follicles, follicular size is a useful indicator of oocyte recovery, fertilization, and cleavage in IVF cycles. For optimal results, the follicular fluid volume in gonadotropin- and hCG-stimulated cycles should be > 1 mL, which corresponds to a follicle diameter of > 12 mm, and not larger than 7 mL (24 mm). For timing of hCG administration, the number of adequate size follicles appears to be more important than the size of the leading follicle(s).

Adult↗
PubMed1989

Bromocriptine inhibition of anesthesia-induced hyperprolactinemia: effect on serum and follicular fluid hormones, oocyte fertilization, and embryo cleavage rates during in vitro fertilization.

Thirty-two patients undergoing in vitro fertilization (IVF) were given bromocriptine either 1 or 12 hours before anesthesia or received no drug to determine what effect suppression of transient, anesthesia-induced hyperprolactinemia would have on peripheral and follicular fluid hormones, fertilization and cleavage rates, and pregnancy. Thirty minutes after anesthesia, there was a 120-ng/mL rise in serum prolactin (PRL) in control patients versus an insignificant change in women given bromocriptine. Levels of PRL in follicular fluid were significantly less, and estradiol (E2) levels were higher (P less than 0.05) in all bromocriptine-treated patients compared with controls, whereas follicular fluid levels of progesterone (P), inhibin activity, and midluteal serum P were unaffected. Although fertilization and pregnancy rates were similar, a greater proportion of fertilized oocytes from bromocriptine-treated patients advanced to cleaving embryos compared with controls (95% versus 63%, respectively; P less than 0.001). We conclude that bromocriptine, given before anesthesia, can suppress transient, anesthesia-induced hyperprolactinemia and dramatically alter follicular fluid concentrations of PRL and E2. Although these changes in hormonal milieu affected neither oocyte fertilization nor pregnancy rate in our IVF patients, they seemed to have a positive influence on embryonic development after IVF.

Adult↗
PubMed1997

Prospective randomized trial of the effect of two flushing media on oocyte collection and fertilization rates after in vitro fertilization.

OBJECTIVE: To assess the value of heparinized saline as a flushing medium for oocyte recovery. DESIGN: Prospective randomized study. SETTING: Academic tertiary referral center for fertility treatment. PATIENT(S): Thirty-five patients, with both ovaries intact having IVF-ET. INTERVENTION(S): Patients were randomized either to have the follicles of the left or right ovary flushed with heparinized normal saline at the time of oocyte recovery for IVF-ET. The contralateral ovary was flushed with heparinized culture medium. Oocytes obtained from each side were cultured separately and assessed for fertilization 18-21 hours after insemination. MAIN OUTCOME MEASURE(S): Collection and fertilization rates. RESULT(S): A total of 481 follicles were aspirated yielding 366 oocytes. Of these, 240 fertilized. From the side flushed with saline 185 oocytes were collected from 237 follicles, which was not significantly different from 181 oocytes collected from 244 follicles on the side flushed with culture medium (odds ratio = 1.23; 95% confidence interval = 0.79-1.92). Similarly, there was no significant difference observed in fertilization rates between oocytes obtained after saline (median 71.4%) and culture medium flush (median 75.0%) (odds ratio = 1.08; 95% confidence interval = 0.68-1.72). CONCLUSION(S): Heparinized normal saline is an equally good but cheaper and more convenient medium than standard heparinized culture medium and could replace it for flushing follicles during oocyte recovery for IVF-ET procedures.

Adult↗
PubMed1999

Prediction of the rates of fertilization, cleavage, and pregnancy success by cumulus-coronal morphology in an in vitro fertilization program.

OBJECTIVE: To examine the relation between the grading of cumulus-coronal morphology at oocyte retrieval and the rates of fertilization, cleavage, and pregnancy success in IVF-ET cycles. DESIGN: Retrospective study. SETTING: University-affiliated medical center. PATIENT(S): Infertile women who underwent IVF-ET treatment. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Fertilization and cleavage of the oocytes and the pregnancy outcome. RESULT(S): Mature grade 3 cumulus-oocyte complexes (COCs) constituted the highest percentage among all grades and had a higher fertilization rate than COCs of other grades (77% versus 65%, 43%, and 28% for grades 2, 1, and 4, respectively). The cleavage and polyspermy rates did not correlate with cumulus-coronal morphology grading. The pregnancy rate was higher in cycles with >50% grade 3 COCs than in cycles with < or =50% grade 3 COCs (32% versus 16%). In cycles with >80% grade 3 COCs, the pregnancy rate was 57%. The correlation between the percentage of grade 3 COCs and the pregnancy rate was independent of patient age and the number of COCs retrieved. CONCLUSION(S): The cumulus-coronal morphology grade correlates with the fertilization rate but not with the cleavage or polyspermy rate. In vitro fertilization cycles that have a greater percentage of grade 3 COCs have an increased chance of resulting in pregnancy. The cumulus-coronal morphology grade predicts pregnancy success in IVF-ET cycles.

Adult↗
PubMed2004

Influence of sperm:oocyte ratio during in vitro fertilization of in vitro matured cumulus-intact pig oocytes on fertilization parameters and embryo development.

The present study was conducted to evaluate the influence of sperm:oocyte ratio during in vitro fertilization (IVF) of in vitro matured cumulus-intact oocytes on fertilization parameters and embryo development in pigs. In vitro matured oocytes surrounded by intact cumulus cells (COC) were inseminated with frozen-thawed spermatozoa at different sperm:oocyte ratios (2000:1, 3000:1, 4000:1, 6000:1, and 8000:1). Denuded oocytes inseminated with 2000 frozen-thawed spermatozoa:oocyte were the control group. A total of 2546 oocytes in five replicates were exposed to spermatozoa for 6 h and then cultured in embryo culture (EC) medium for 6 h (pronuclear formation) or 7 days (blastocyst formation: BF). The penetration rate increased in the COC groups with the sperm:oocyte ratio, reaching the highest rates with 8000:1 spermatozoa:oocyte (72.1 +/- 6.5%), similar to the control (73.5 +/- 3.5%). However, the monospermy was highest with the lower spermatozoa:oocyte rates (82.6-94.8%) and decreased drastically (P<0.05) in the COC group fertilized with 8000 sperm:oocyte (36%). The efficiency of fertilization (number of monospermic oocytes/total number of inseminated oocytes) showed no difference among the COC groups (20-30%) but they were significantly lower (P<0.007) than those obtained by the control group (43.7 +/- 2%). Embryo development was highest in the control group (58% for cleavage and 23% for BF) but not significantly different with the 6000 and 8000 sperm:oocyte COC groups (47 and 50% for cleavage and 19 and 17% for BF, respectively). These results indicate that the use of COC for IVF involves a drop in the efficiency of the fertilization and the necessity to increase the frozen-thawed sperm:oocyte ratio three to four times more to obtain similar embryo development to denuded oocytes.

Animals↗
PubMed2003

Serum prolactin and TSH in an in vitro fertilization population: is there a link between fertilization and thyroid function?

PURPOSE: Measurements of TSH and prolactin are generally included in the evaluation of female infertility, but their value in women coming to in vitro fertilization (IVF) has been questioned. METHODS: In this study, we sought to investigate whether prolactin or TSH, measured in 509 specimens collected prior to therapy, predicted outcome in a prospective study of couples undergoing IVF between 1994 and 2001. RESULTS: TSH was higher in women whose fertility problem was attributed to a male factor, and prolactin was lower if the measurement was taken during menses. TSH and prolactin were positively correlated (p < 0.0001). Neither TSH nor prolactin levels correlated with overall IVF outcome; however, TSH levels were significantly higher among women who produced oocytes that failed to be fertilized and this finding persisted after adjustment for several covariates, including sperm motility. Among women who had a least one oocyte inseminated, the likelihood that they would have fewer than 50% of their eggs fertilized was significantly related to higher TSH levels in a multivariate model. CONCLUSION: We conclude that TSH may predict poor fertilization in IVF and reflect the importance of thyroid hormones in oocyte physiology.

Female↗
PubMed1997

Male-dependent variability of fertilization and embryo development in two bovine in vitro fertilization systems and the effects of casein phosphopeptides (CPPs).

This study investigated the effects of semen from five different bulls and two different ejaculates of the same bull on penetration, cleavage, blastocyst formation, and cell allocation in bovine blastocysts produced in vitro. Casein phosphopeptides (CPPs) were tested for their ability to enhance fertilization and minimize variability among bulls and ejaculates. In Experiment 1, the BO-fertilization system was employed. Penetration and polyspermy both displayed great variation among bulls and between ejaculates, whereas no significant differences were observed in cleavage and blastocyst-formation rates. Similar variability was observed in penetration, polyspermy, cleavage, blastocyst-formation rates and cell allocation and distribution when the two fertilization systems, TALP and BO, were compared in Experiment 2. The BO-system supported penetration and polyspermy better (P < 0.05) than the TALP-system, whereas the TALP-system was superior (P < 0.05) in supporting cleavage and blastocyst formation. Significant interactions existed between bulls and the fertilization system employed. It is concluded that the success of in vitro fertilization is markedly dependent on individual bulls as well as on ejaculates from the same bull. CPPs are able to enhance penetration and embryo development in certain bulls or ejaculates and thus contribute to reducing the degree of individual variability, but they do not generally improve the success of bovine embryo production in vitro.

Animals↗
PubMed1997

Fertility and contraception in Europe: the case of low fertility in southern Europe.

Demographic trends in Europe can be summarized in terms of a declining proportion of its population in the world total, low fertility, and a slow population growth. Fertility in Western Europe was already below the replacement level in 1970-75 and has remained low. Fertility has substantially declined in Eastern and Southern Europe, with Italy and Spain recording one of the lowest levels (1.2 children per woman) in 1994. Some explanations of the dramatic fertility decline in Southern Europe are: (1) The emancipation of women and their increased participation in the labor force; (2) Economic aspects such as costs for child care and education; and (3) The couple's motivation for low fertility because of the expanded choices for travel and leisure and their concerns for improving their standard of living. Social pressures on childbearing outside marriage remain quite strong; cohabitation and extramarital births is Southern Europe are not as prevalent as in other European regions and there is a trend toward delaying the birth of the first child rather than foregoing childbearing.

Adult↗
PubMed1997

Relationship of human follicular diameter with oocyte fertilization and development after in-vitro fertilization or intracytoplasmic sperm injection.

The aim of this work was to evaluate the relationship between follicular size at the time of oocyte retrieval, and the subsequent oocyte competence to be fertilized and to develop in vitro. All the obtained oocytes were classified according to the corresponding volume of aspirated follicular fluid. Aspirated volume of follicular fluid <2 ml corresponded to a follicular diameter <16 mm and constituted the small size group. Volume of follicular fluid from 2 to 6 ml corresponded to a diameter from 16 to 23 mm and constituted the medium size group. The large size group contained follicles with diameter >23 mm and corresponded to an aspirated volume of follicular fluid of >6 ml. A progressive and significant increase in the rates of oocytes with a first polar body was observed from the small size group to the other groups and from the medium to the large size group: 75.3, 85.9 and 95.3% respectively. After classical in-vitro fertilization (IVF), significantly better rates of fertilization and development were obtained in the medium size group compared to the two other groups. Moreover, a positive relationship was observed between follicular diameter and rates of embryos scored as 'good' when oocytes were fertilized by intracytoplasmic sperm injection (ICSI). These results demonstrated that follicular size is positively related to the oocyte ability to be fertilized and to develop. Although oocytes from small follicles gave lower percentages of development probably due to partial oocyte incompetence, they allowed an increase in the total number of embryos scored as 'good'.

Cell Size↗
PubMed1998

Case report: high fertilization rate in conventional in-vitro fertilization utilizing spermatozoa from an oligozoospermic subject presenting microdeletions of the Y chromosome long arm.

A case is reported in which a high fertilization rate was achieved by conventional in-vitro fertilization (IVF), using spermatozoa from an oligozoospermic man carrying a microdeletion of the long arm of the Y chromosome. The patient presented with idiopathic infertility of 10 years duration; the fertility status of his wife was completely normal. After IVF, five out of eight oocytes retrieved showed normal fertilization and four showed normal embryo cleavage. Four embryos were transferred; however, pregnancy did not result. These results demonstrate that spermatozoa from oligozoospermic patients carrying a Yq microdeletion are fully competent in achieving capacitation, acrosome reaction and fertilizing ability during IVF. Therefore, although definitive conclusions cannot be made from a single case report, we suggest that Yq microdeletion analysis should be considered in oligozoospermic patients undergoing conventional IVF.

Adult↗
PubMed1983

A study of factors affecting the success of human fertilization in vitro. II. Influence of semen quality and oocyte maturity on fertilization and cleavage.

The incidence of in vitro fertilization was analyzed with respect to the degree of cumulus dissociation (expansion) at the time of oocyte recovery and also the semen quality. Of the oocytes surrounded by perfectly ("++") or moderately ("+") dissociated cumuli, 78.6% and 30.8%, respectively (P less than 0.001), were fertilized when the husband's semen analysis was in the normal range. The proportion of fertilized oocytes was not decreased in cases of polyzoospermia (greater than 130 X 10(6) spermatozoa/ml), but was decreased (P less than 0.05) when the semen analysis revealed other anomalies: oligozoospermia (less than 15 X 10(6) spermatozoa/ml), asthenozoospermia (less than 50% motile cells) or teratozoospermia (greater than 50% abnormal spermatozoa). The proportion of fertilized eggs cleaving in vitro was unaffected by semen quality but was lower when "+" cumulus oocytes were collected than when "++" cumulus oocytes were obtained (58.3% vs. 87.0%, P less than 0.02). In vitro incubation of the oocyte prior to insemination increased the incidence of fertilization by about 28% for both "+" (22.2 to 50.0%) and "++" (65.7 to 93.9%) cumulus oocytes. Finally, 67.6% of "++" cumulus oocytes developed into embryos when the insemination with spermatozoa from normal semen samples was delayed by several hours, compared with only 29.0% when the conditions were suboptimal ("+" cumulus oocyte, abnormal semen analysis or no delay prior to insemination). Eight pregnancies began following the replacement of 38 embryos in 34 patients. Six spontaneous abortions occurred, and chromosomal abnormalities were proven in the two cases analyzed. Two pregnancies continued for more than 3 months, resulting in term deliveries of two normal babies.

Cell Division↗
PubMed2002

Fertilization of mouse oocytes from in vitro-matured preantral follicles using classical in vitro fertilization or intracytoplasmic sperm injection.

Early preantral mouse follicles with a diameter of 110-160 microm were cultured in vitro for 10 or 12 days. Mature oocytes were retrieved following hCG, and fertilization was attempted either by in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Two-cell and blastocyst formation rates and blastocyst cell numbers were compared between 10-day and 12-day in vitro-matured oocytes versus in vivo-matured oocytes. Uncleaved IVF oocytes were subjected to chromosome analysis. The 2-cell formation rate was significantly improved by ICSI compared with IVF both in 10-day (72.1% versus 56.1%; P = 0.03) and 12-day cultures (74.1% versus 54.5%; P = 0.028). Cytogenetic analysis of uncleaved MII oocytes following IVF showed that about 30% of MII oocytes showed no sign of sperm penetration. The blastocyst formation rate was significantly lower in 12-day versus 10-day cultures, whether fertilization was by IVF (40.7% versus 62.4%, P = 0.016) or by ICSI (32.5% versus 57.1%, P = 0.035). Blastocyst cell numbers from IVF and ICSI 10-day groups were similar and both significantly higher (P < 0.001) than from IVF 12-day cultures. All above expressed values were significantly higher for in vivo-matured oocytes. In conclusion, fertilization of oocytes from in vitro-matured mouse preantral follicles can be optimized with ICSI, giving significantly higher 2-cell formation rates than IVF. Blastocyst formation rate was not influenced by the technique of fertilization but rather by the extent of the in vitro culture period. Best results on preimplantation development of oocytes for in vitro-matured preantral follicles were obtained with ICSI on oocytes from 10-day in vitro cultures.

Animals↗