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Successful fertilization, embryo development, and pregnancy in human in vitro fertilization (IVF) using a chemically defined culture medium containing no protein.

A randomized control trial involving the fertilization and culture of human embryos in culture medium (T6) containing either 10% maternal serum or no protein or amino acid supplement was carried out to assess the effect of deletion from culture of all fixed nitrogen on fertilization, embryo development, and embryo viability. There was no difference in fertilization rates (68 vs 69%), development of apparently normal embryos (96 vs 97%), pregnancy rate (18 vs 14%), or birth rate (13 vs 11%) between protein-containing and protein-free media. Deletion of protein from the culture medium may enable the constitution of more appropriate and defined culture media for human in vitro fertilization (IVF).

Culture Media↗

The prognostic importance of the number of oocytes retrieved and estradiol levels in poor and normal responders in in vitro fertilization (IVF) treatment.

A low response to ovarian stimulation in in vitro fertilization poses a unique therapeutic challenge. Gonadotropin-releasing hormone agonists (GnRHa) have been suggested as a modality for treatment of this condition. In this study, we analyzed the results of 880 in vitro fertilization treatment cycles with respect to modality of ovarian stimulation, degree of hormonal response, and number of oocytes retrieved. In patients with estradiol (E2) levels less than 501 pg/ml on the day of human chorionic gonadotropin administration, 27% pregnancy rate was achieved with clomiphene citrate (CC) combined with human menopausal gonadotropin (hMG), compared to 15.1% (P less than 0.005) with hMG alone and 20.8% (NS) with GnRHa and hMG. Pregnancy rates were not lower in these patients compared to patients with higher estradiol levels in the different stimulation protocols, but pregnancy rates were significantly lower in cycles during which three or fewer oocytes were retrieved, compared to those in which four or more oocytes were retrieved (10.8 vs 23.8%; P less than 0.0005). In low-retrieval cycles pregnancy rates actually decreased with increasing levels of estradiol. Our results indicate that the number of oocytes retrieved is a better prognostic parameter than E2 levels in predicting the outcome of in vitro fertilization treatment and that GnRHa in the long protocol do not seem to be superior to CC combined with hMG for the treatment of poor responders.

Biomarkers↗

Serum and follicular fluid (FF) estradiol (E2) levels in ovarian hyperstimulation syndrome (OHSS) during in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) conception cycles after pituitary suppression.

Initial hope that ovarian hyperstimulation syndrome (OHSS) would be less likely to occur after pituitary suppression with gonadotropin releasing-hormone agonists (GnRH-a) has not been substantiated. GnRH-a/human menopausal gonadotropin (hMG) protocols often lead to OHSS with markedly elevated circulating estradiol (E2) levels in susceptible patients. This study was undertaken to determine whether or not intrafollicular E2 secretion is increased in these cases. Fifty-two in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) conception cycles treated with GnRH-a/hMG were included in the study. GnRH-a, leuprolide, 0.5 mg, was administered subcutaneously from day 20 of the preceding cycle and the ovaries were stimulated with hMG, 75-225 IU bid intramuscularly, followed by human chorionic gonadotropin (hCG), 5000 IU. Twenty cycles (Group I) were associated with moderate or severe OHSS and 32 cycles (Group II) did not result in OHSS. E2 was measured in the serum on the day of hCG (day 0), on the day of oocyte retrieval (day 2), and at midluteal phase (days 6-8), as well as in the follicular fluid (FF) using a solid-phase direct RIA. Mean serum E2 was significantly higher at all three sampling times in Group I (OHSS) than in Group II. Both the number of follicles and the number of oocytes were also significantly higher in Group I.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effect of chlamydial antibodies on the outcome of in vitro fertilization (IVF) treatment.

OBJECTIVE: Our objective was to study the relationship between the presence of Chlamydia trachomatis antibodies and the success of IVF treatment. DESIGN: We evaluated 118 in vitro fertilization and embryo transfer (IVF-ET) treatment cycles from 51 couples with a history of infertility lasting for at least 2 years. All women starting a treatment cycle had their serum chlamydial antibody titers measured by indirect immunofluorescent technique. All patients received similar ovarian stimulation regimens and the oocytes collected were inseminated with similar concentrations of motile sperm. Clinical data from couples where the female partner had C. trachomatis ab titers > or = 40 have been compared with the equivalent data from couples where the female partner had C. trachomatis ab titers < 40. RESULTS: There was no statistically significant difference between the two groups concerning age, infertility period, oocytes collected, oocytes fertilized, and fertilization rate, and the pregnancy rates were comparable. CONCLUSION: Previous exposure to C. trachomatis did not alter the success rate of IVF-ET.

Adult↗

In-vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT).

In the past decade, the use of in-vitro fertilization and embryo transfer (IVF-ET) procedures as a means of treatment of infertility has become a clinical reality. More recently, similar yet differing techniques have been described that further expand options of treatment of infertile couples. This article summarizes recent advances in IVF and compares it with the most widely recognized of these alternatives, gamete intrafallopian transfer (GIFT). In addition to comparing and contrasting these new technologies, the authors attempt to highlight unresolved questions about their use.

Fallopian Tubes↗

Luteinizing response to human chorionic gonadotropin does not predict outcome in gonadotropin releasing hormone agonist-suppressed/human menopausal gonadotropin-stimulated in vitro fertilization (IVF) cycles.

OBJECTIVE: The purpose of this study was to determine if early luteinizing potential in gonadotropin releasing hormone agonist (GnRH-a)-suppressed/human menopausal gonadotropin (hMG)-stimulated IVF cycles is predictive of cycle outcome. DESIGN, PATIENTS: The study was a prospective evaluation of 41 women beginning a GnRH-a-suppressed/hMG-stimulated IVF cycle. SETTING: The in vitro fertilization program of a tertiary care institution was the study setting. MAIN OUTCOME MEASURES: The main outcome measures were (1) estradiol (E2) and progesterone (P) levels on the day of human chorionic gonadotropin (hCG) administration and the following day and (2) the ovarian response to ovulation induction and clinical outcome. RESULTS: Ten of the 41 women achieved a clinical pregnancy (24.4%). There was no significant difference in progesterone (P) levels on the day of or the day following hCG administration between the pregnant and the nonpregnant groups. Both groups exhibited a significant rise in P level in response to hCG. There was no significant difference in E2 levels on the day of hCG between the two groups. The serum E2 did not rise significantly in response to hCG in either group. Patients who became pregnant had significantly more oocytes retrieved, fertilized, cleaved, and transferred. CONCLUSIONS: Clinical response and outcome in GnRH-a-suppressed/hMG-stimulated IVF cycles are not predicted by early luteinizing potential as indicated by the response of E2 or P to hCG.

Adult↗

The development of an efficient ambulatory in vitro fertilization (IVF) and embryo transfer (ET) program using ultrasonically guided oocyte retrieval.

Until recently most of the current in vitro fertilization and embryo transfer (IVF-ET) programs used laparoscopy for oocyte retrieval and included hospitalization in the treatment. The establishment of a successful ambulatory IVF program that uses ultrasonically guided oocyte aspiration, is described. Between August 1985 and February 1987, 384 percutaneous transvesical ultrasonically and transvaginally guided follicle aspirations were performed on 414 patients, following ovarian stimulation with hMG and hCG. Ninety two clinical pregnancies were confirmed by ultrasound. The clinical pregnancy rate was 26.8% when based on the number of embryo transfers. There were 16 multiple pregnancies (17.4%), 18 miscarriages (19.5%), and two tubal pregnancies (2.1%). The most important factors contributing toward the success of this program were rigid adherence to clinical and laboratory protocols, and the maintenance of a strict quality control. The ambulatory management was very well accepted by the patients, who during the entire period of ovarian stimulation had minimal disruption of their routine activities.

Adult↗

In vitro DNA fluorescence after in vitro fertilization (IVF) failure.

PURPOSE: A pilot study was performed to test the diagnostic value of in vitro DNA fluorescence in oocytes that failed to fertilize after IVF. Ten patients with a cleavage rate less than 20% after IVF were included. RESULTS: Uncleaved oocytes were observed by fluorescence microscopy after incubation with the DNA fluorescent dye Hoeschst 33342. Four main causes which may have contributed to the low cleavage rate were found: (1) sperm incapacity to penetrate the oocyte despite the absence of the usual criteria for male infertility, (2) oocyte immaturity, (3) delayed fertilization, and (4) oocyte abnormalities revealed by aberrations in the morphology of the female chromatin. CONCLUSIONS: The possibility of a rapid and detailed analysis of the maturational status of unfertilized oocytes, the morphology of the female chromatin, the presence and quantity of spermatozoa tightly bound to the zona pellucida, and sperm penetration into the oocyte without subsequent pronucleus formation, using DNA fluorescence, allows us to clarify further the cause of fertilization failure and to orient infertility treatment toward the male, the female, or both partners.

Adult↗

Subzonal insemination (SUZI) or in vitro fertilization (IVF) in microdroplets for the treatment of male-factor infertility.

PURPOSE: The results of subzonal insemination (SUZI) and in vitro fertilization with microdroplet insemination used in couples with male-factor infertility are presented. RESULTS: The total fertilization rate was 17.4% for SUZI (n = 89) and 49.3% for microdroplet IVF (n = 100). The fertilization rate for standard IVF (n = 510), not including any male-factor infertility and performed during the same period, was 73.2%. The "take-home baby rate" per started cycle and per embryo transfer (ET), respectively, was 10 and 17.6% for SUZI and 20 and 24.7% for microdrop IVF. For standard IVF these figures were 27 and 31.7%. CONCLUSION: It was concluded that microdroplet IVF can be used with good results in cases of moderate male-factor infertility. The normal (2PN) fertilization rate with the SUZI technique was only 15.1%. However, despite the low fertilization rate, SUZI should be considered when dealing with severe male-factor infertility.

Fertilization in Vitro↗

Maternal/paternal sharing of DQ-Alpha type II histocompatibility antigens not associated with pregnancy outcome following in vitro fertilization (IVF)-embryo transfer (ET).

PURPOSE: To determine if maternal/paternal sharing of DQ alpha major histocompatibility (MHC) type II antigens is associated with reduced pregnancy and implantation rates following in vitro fertilization-embryo transfer (IVF-ET). METHODS: Prospective study with type II MHC DQ alpha alleles detected by polymerase chain reaction (PCR) technology using Perkin Elmer Amyli-type HLA DQ alpha PCR amplification and typing kit. The tests were only performed on patients having their first IVF cycle. RESULTS: No difference was found in clinical pregnancy rates per transfer between those couples sharing DQ alpha I alleles and those who did not (43.7% vs 40%). There were no spontaneous abortions in the group sharing DQ alpha I alleles. CONCLUSION: Maternal/paternal sharing of DQ alpha I antigens does not reduce fecundity following IVF-ET.

Adult↗

The diagnostic value of seminal adenosine triphosphate (ATP) in an in vitro fertilization (IVF) program.

The level of adenosine triphosphate (ATP) was quantitated in semen samples used for in vitro fertilization of human oocytes. Seminal ATP level correlated with the concentration and percentage motility of spermatozoa but not with the in vitro fertilization rate of human oocytes. Seminal ATP measurement appears to have little diagnostic value in predicting the fertilizing capacity of spermatozoa as evaluated by the multivariate stepwise discriminant analysis.

Adenosine Triphosphate↗

A prospective study of the effects of female and male marijuana use on in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) outcomes.

OBJECTIVE: This study was undertaken to examine whether marijuana use affects in vitro fertilization and gamete intrafallopian transfer (IVF/GIFT). STUDY DESIGN: Prospective study of 221 IVF/GIFT couples. RESULTS: Amount of lifetime heavy marijuana use adversely affected IVF/GIFT. Women smoking more than 90 times in their lifetime had 27% fewer oocytes retrieved (P = .03) and 1 fewer embryo transferred (P < .05). Women smoking marijuana more than 10 times in their lifetime had infants 17% (P = .01) smaller at birth. If men smoked marijuana 11 to 90 times in their lifetime, there was a 15% decrease in infant birth weight (P = .03); if this increased to more than 90 times, there was a 23% decrease (P = .01). Timing also played a role. Women smoking marijuana 1 year before IVF/GIFT had 25% fewer oocytes retrieved (P = .03), whereas couples had 28% (P = .04) fewer oocytes fertilized. Women and men who smoked in the past 15 years, had 12% (P = .04) and 16% (P = .03) smaller infants, respectively. CONCLUSION: Both timing and amount of marijuana use negatively affected IVF/GIFT.

Adult↗

Developmental assessment of twenty in vitro fertilization (IVF) infants at their first birthday.

The pregnancy details, delivery outcome, and developmental status as measured on the Griffiths Developmental Scales are provided on the first 20 infants reaching their first birthday following in vitro fertilization-embryo transfer (IVF-ET) within the PIVET Programme. An increased rate of preterm delivery, intrauterine growth retardation, and cesarean sections was noted. One significant and two minor abnormalities were detected and only one infant was slightly under the expected developmental assessment at 1 year on the corrected general quotient of the Griffiths Developmental Scales for children.

Child Development↗

Comparison between the hypoosmotic swelling test and morphology evaluation using strict criteria in predicting in vitro fertilization (IVF).

BACKGROUND: The role of male factor with respect to sperm morphology, progressive motility, and density is studied under in vitro conditions. METHODS: The semen samples of 67 males participating in an in vitro fertilization program were evaluated by the conventional WHO criteria of spermatogram, by morphology evaluation using strict criteria (MEUSC), and by the hypoosmotic swelling test (HOST). All sperm tests were performed in the original semen sample as delivered on the day of IVF, before further sperm treatment. The correlations between these parameters and the fertilization outcome were evaluated and their predictive values were calculated. RESULTS: When the patients were divided into two groups, namely, fertile (fertilization rate per oocyte greater than 0%) and infertile (fertilization rate per oocyte = 0%), only mean sperm density and morphology were significantly different between the groups (P less than 0.05). The correlation with fertilization rate in vitro was in favor of MEUSC. CONCLUSIONS: Our results show that the HOST is inferior to MEUSC and conventional WHO sperm analysis in predicting fertilization in vitro.

Cell Membrane↗

In vitro fertilization (IVF)-surrogacy: application of IVF to women without functional uteri.

Women with absent or dysfunctional uteri consented to controlled ovarian stimulation, ovum aspiration, in vitro fertilization, and embryo culture. Cleaving preembryos were transferred to recipient (surrogate) women whose menstrual cycles were in approximate synchrony with the ovum donor. None of the embryo recipients received medication. Six cases are described, resulting in one spontaneous loss at 6 weeks, four full-term deliveries, and one ongoing pregnancy. HLA typing demonstrated all babies to be genotypic offspring of the gamete donors.

Female↗

Spontaneous conception in a 50-year old woman after giving up in-vitro-fertilization (IVF) treatments: involvement of the psychological relief in successful pregnancy.

A healthy woman, married at age 45, hoped to have their child without delay. She and her husband subsequently decided to pursue in-vitro-fertilization and embryo transfer (IVF-ET). In the succeeding year, ET was performed twice from four oocyte retrieval attempts, which represented six ovarian stimulation cycles. Pregnancy was not achieved. Because of her poor responses to ovarian stimulation, inferior oocyte grading and prohibition of donor oocyte usage in Japan, the couple decided to discontinue further IVF treatment at age 48 years, 10 months. One and one-half years later, at age 50 years, 3 months she presented to our clinic eight weeks pregnant. At term she delivered vaginally a 2740 g healthy infant at 38 weeks gestation; Apgar scores were 9 and 10. During her pregnancy, she willingly participated in our questionnaire designed to provide information about her psychological well-being during the past three years. During the time spanning her treatment for infertility, anxiety, depression, irritability, fatigue and grief were revealed to coexist with her high hopes of having a child. After termination of infertility treatments these adverse psychological findings were markedly lessened and her vigour was restored. Stopping infertility treatment might be a viable alternative for achieving pregnancy in similarly psychologically-challenged infertile women.

Embryo Transfer↗

Ovarian stimulation with gonadotropin-releasing hormone (GnRH) analogue improves the in vitro fertilization (IVF) pregnancy rate with both transvaginal and laparoscopic oocyte recovery.

The relative impact of ovarian stimulation protocol and oocyte retrieval technique on success rates of in vitro fertilization program was studied in 200 patients. Sixty-three patients received gonadotropin-releasing hormone analogue (GnRHa) with human menopausal gonadotropin (hMG), and 137 received hMG only. The GnRHa + hMG protocol resulted in higher pregnancy rates than the hMG-only protocol (19.0 vs 9.5%, respectively; P less than 0.01) despite a lower cleavage rate. Oocyte retrieval was performed via laparoscopy in 100 patients and transvaginally in 100 patients. The number of oocytes recovered per cycle was 6.1 +/- 3.9 with laparoscopy and 7.0 +/- 3.1 transvaginally. Pregnancy rates were similar for both retrieval techniques (13 and 12%, respectively). A breakdown of these results showed that the advantage for the GnRHa + hMG protocol was not affected by the oocyte retrieval technique. A comparison of simultaneous blood and follicular fluid pH measured every 10 min during laparoscopy and transvaginal oocyte recovery revealed a constant decline in follicular fluid pH during laparoscopy, while no changes were observed during the vaginal procedure. We conclude that the improvement in in vitro fertilization results during the period of our study is due primarily to the introduction of GnRHa + hMG protocol rather than the method of oocyte retrieval.

Biopsy, Needle↗

[Effect of various factors on in vitro fertilization (IVF) and embryo transfer in the human in clomiphene/hMG/hCG stimulated cycles].

In the present study we tried to analyse data obtained on the occasion of an in vitro fertilization programme at the department of obstetrics and gynecology of the University of Kiel, to extract some of the important factors, because their positive or negative influence on the success of extracorporal insemination, embryo transfer and implantation has not been clearely defined till now and to value their concern. Between July 1982 and February 1983 105 sterile patients with ovulatory menstrual cycles have been treated by the technique of in vitro fertilization and embryo transfer, successively. Follicular growth was stimulated by 150 mg clomiphen daily from 3rd to 7th day and application of 225 IU hMG on 6th, 8th and 10th day. Finally ovulation was timed 36 hours before follicular puncture by 5000 IU hCG. In 24 per cent endogenous LH peak could be observed. In 86 per cent of the 80 patients exactly timed one oocyte was to be found minimally. In 68 per cent of these patients after all an embryo transfer was done (24 times one embryo, 13 times two, 9 times three and once four embryos). 89% of the embryos have been transfered as 4-or 8-cell stage, only 11 per cent as two cell stage. Transfer took place 48 to 50 hours following insemination. There was no correlation between the maturity classified morphologically of the oocytes and the diameter of their follicles. The speed of development of the embryos was not correlated with the size of their follicles. too. In 60-70 per cent of all cases, in which an embryo transfer was possible, husbands had normal spermiogram. The percentage of unfertilised oocytes following insemination using non-optimal sperma was increased in all types of cycles, additionally there was a reduction of the developing speed of resulting embryos. Doubtless two the four pregnancies resulted after insemination using sperma of less quality. The conjugation rate is said to be not correlated directly with the exactness of timing of the application of hCG with reference to follicular maturity, but in all pregnancies hCG was injected simultaneously with the peak of estradiol.

Chorionic Gonadotropin↗