Women with declining ovarian reserve may demonstrate a decrease in day 3 serum inhibin B before a rise in day 3 follicle-stimulating hormone.

OBJECTIVE: To test the hypothesis that women with declining ovarian reserve may demonstrate a decrease in day 3 serum inhibin B levels before a rise in day 3 serum FSH levels. DESIGN: Case-control study. SETTING: Tertiary care fertility center. PATIENT(S): One hundred nine women with nonovarian infertility (tubal factor or male factor) and 47 women with declining ovarian reserve who underwent assisted reproductive techniques. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Serum inhibin B and FSH levels, number of ampules of gonadotropins administered, E2 levels on the day of hCG administration, number of oocytes retrieved, clinical pregnancy rate, and cycle cancellation rate. RESULT(S): Women who had declining ovarian reserve as demonstrated by an increased gonadotropin requirement, a decreased E2 response, fewer retrieved oocytes, a lower clinical pregnancy rate, and a higher cycle cancellation rate had lower day 3 serum inhibin B levels despite having nonelevated day 3 FSH levels similar to those of women with nonovarian infertility. CONCLUSION(S): Women with declining ovarian responsiveness and clinical outcomes consistent with declining ovarian reserve had decreased day 3 serum inhibin B levels despite having nonelevated day 3 serum FSH concentrations. Declining ovarian reserve may be demonstrated by a decrease in day 3 inhibin B levels before a rise in day 3 FSH levels.

Influence of endogenous progesterone on alprazolam pharmacodynamics.

The results of a recently completed study demonstrated that postmenopausal women were more sensitive to triazolam-induced psychomotor performance impairment when progesterone was administered concomitantly. That clinical evidence agrees with the emerging in vitro information regarding the rapid membrane effects of a progesterone metabolite that positively modulates the gamma-aminobutyric acid-A-benzodiazepine receptor complex. The objective of this study in premenopausal women was to determine whether the response to a benzodiazepine is altered when endogenous progesterone concentrations are high (luteal phases of a menstrual cycle) compared with when progesterone concentrations are low (follicular phases of a menstrual cycle). The pharmacokinetics and pharmacodynamics of oral alprazolam were evaluated in twelve healthy, normally menstruating women who were not receiving oral contraceptive agents. On two separate occasions, once during each phase of the menstrual cycle, the women randomly received an oral alprazolam 2-mg dose. Blood samples were collected, and psychomotor performance tests were conducted at selected times before and after dosing. These data show that fluctuations of endogenous progesterone across the menstrual cycle do not influence alprazolam pharmacodynamics. Despite endogenous progesterone concentrations being significantly higher during the midluteal than during the midfollicular drug administrations, no differences were observed in either the digit-symbol substitution test, card sorting by suit, or sedation scores on these two occasions. No pharmacokinetic differences were observed between the two menstrual cycle-phase drug administrations. In conclusion, the lack of changes during the menstrual cycle in demonstrable cognitive impairment and pharmacokinetics after alprazolam administration is reassuring. This implies that a dose adjustment made on the basis of menstrual timing is not required.

Effects of ovarian source, patient age, and menstrual cycle phase on in vitro maturation of immature human oocytes.

OBJECTIVE: To determine the efficiency of in vitro maturation, expressed by nuclear maturation, of oocytes aspirated during gynecologic surgeries or collected from excised ovaries. To assess the effect of patient age and cycle phase at collection on the oocyte's ability to mature in vitro. To examine the time course of oocyte maturation in vitro. DESIGN: Nuclear maturation based on patient criteria compared. SETTING: University-based IVF program and research center. PATIENT(S): Consented patients undergoing gynecologic surgeries or patients undergoing oophorectomy. INTERVENTION(S): Oocytes were maintained in culture for 48 hours and evaluated for maturation. MAIN OUTCOME MEASURE(S): Nuclear maturation evaluated as germinal vesicle breakdown (GVBD) or progression to the metaphase II (MII) stage. RESULT(S): A significantly higher percentage of oocytes collected during the follicular phase of the menstrual cycle underwent GVBD than did oocytes collected during the luteal phase (60% versus 48%, respectively). The percentage of oocytes reaching the MII stage, from these two groups, was not different. No statistically significant differences in maturation were observed in oocytes from different ovarian sources or from patients >40 or <40 years of age. CONCLUSION(S): These data suggest that oocytes collected during the follicular phase are more likely to undergo GVBD than oocytes collected during the luteal phase. In this study, ovarian source, age, or cycle phase did not influence the final meiotic maturation of oocytes to metaphase II.

Activin A stimulates meiotic maturation of human oocytes and modulates granulosa cell steroidogenesis in vitro.

OBJECTIVE: To test the hypothesis that activin A promotes in vitro human oocyte meiotic maturation while inhibiting steroid secretion by nonluteinized antral granulosa cells. DESIGN: Prospective randomized controlled study. SETTING: A university medical center. PATIENT(S): Nine women ranging in age from 31-44 years who were undergoing oophorectomy for nonovarian pathology. INTERVENTION(S): Analysis of meiotic maturation of oocytes and steroid secretion by granulosa cells cultured in the presence or absence of activin A. MAIN OUTCOME MEASURE(S): Germinal vesicle breakdown (GVBD) and attainment of metaphase II (MII) in oocytes, and progesterone and E2 secretion by granulosa cells. RESULT(S): Activin A significantly enhanced GVBD (91% vs. 65%) for control and maturation to MII (56% vs. 35% for control) of immature oocytes. Activin A significantly suppressed basal, and inhibin A-and FSH-stimulated progesterone and E2 secretion by nonluteinized granulosa cells. CONCLUSION(S): Activin A is a promoter of oocyte maturation in vitro and a modulator of granulosa cell steroidogenesis in culture.

Elevated level of follicular fluid vascular endothelial growth factor is a marker of diminished pregnancy potential.

OBJECTIVE: To evaluate whether differences in follicular fluid vascular endothelial growth factor (FF VEGF) concentrations are observed between women achieving a clinical pregnancy and those failing to conceive. DESIGN: Retrospective chart review and analysis of FF VEGF concentrations. SETTING: University teaching center. PATIENT(S): Fifty-seven women < or =42 years of age undergoing follicular aspiration in preparation for IVF or GIFT. INTERVENTION(S): Analysis of FF VEGF concentrations and chart review of a single IVF or GIFT cycle. MAIN OUTCOME MEASURE(S): Follicular fluid VEGF concentrations, clinical pregnancy rate, age, ampules of gonadotropins used, oocytes retrieved, peak estradiol serum concentrations, day 3 FSH levels, and fertilization rate. RESULT(S): Women who did not conceive had higher FF VEGF concentrations than women achieving a clinical pregnancy (4.409 + 2,387 versus 2.793 +/- 1,180 pg/mL: P < .001). A negative correlation was observed between FF VEGF concentrations and peak estradiol levels and number of oocytes retrieved. A positive correlation was found for FF VEGF and patient's age and ampules of gonadotropins used. CONCLUSION(S): Elevated FF VEGF concentrations are associated with poor conception rates after IVF or GIFT.

Dimeric inhibin: a direct marker of ovarian aging.

OBJECTIVE: To investigate whether luteal secretion of inhibin-a is altered in the perimenopausal transition and to evaluate whether luteal inhibin secretion is correlated with other markers of ovarian reserve such as FSH and inhibin-b. DESIGN: Prospective study. SETTING: Reproductive Endocrinology Laboratories at The Ohio State University. PATIENT(S): Twenty-five women 39-52 years of age with regular menstrual cycles. INTERVENTION(S): Daily urine samples were monitored (LH predictor kit) to identify the day of ovulation. Blood samples obtained on days 6 and 8 after the LH surge and on day 3 of the subsequent follicular phase were assayed for FSH, E2, progesterone. inhibin-a, and inhibin-b. MAIN OUTCOME MEASURE(S): Serum levels of inhibin-a, inhibin-b, FSH, E2, and progesterone. RESULT(S): Luteal phase inhibin-a and follicular phase inhibin-b were correlated inversely with age in perimenopausal women. In addition, luteal phase inhibin-a and follicular phase inhibin-b levels were correlated inversely with follicular phase FSH levels. CONCLUSION(S): Both luteal phase inhibin-a and follicular phase inhibin-b levels are correlated inversely with age during the fifth decade of life. These findings suggest that corpus luteum function is altered during the perimenopausal transition. Moreover, these direct measures of ovarian function may be more sensitive indicators of "ovarian reserve" than indirect indicators such as pituitary FSH secretion.

Follicular fluid vascular endothelial growth factor concentrations are elevated in women of advanced reproductive age undergoing ovulation induction.

OBJECTIVE(S): To determine whether follicular fluid (FF) from women of advanced reproductive age had a relative deficiency of the angiogenic cytokine vascular endothelial growth factor/vascular permeability factor. Furthermore, we sought to determine whether luteinized granulosa cells secrete vascular endothelial growth factor/vascular permeability factor in response to hypoxia. DESIGN: Retrospective cohort study. SETTING: University teaching hospital. PATIENTS: Women undergoing follicular aspiration after superovulation in preparation for IVF-ET. Women of advanced reproductive age consisted of 21 women > or = 38 years old (range, 38 to 46 years); 15 subjects < or = 30 years served as the control population. INTERVENTION(S): Granulosa cells and FF were collected by transvaginal aspiration 35 hours after hCG. Granulosa cells from two women were cultured for 24 and 48 hours in M199 + 10% fetal bovine serum in 1% O2-5% CO2-94% N2 (hypoxic) or 95% air-5% CO2 (normoxic) without or with 0.1 mol/L cobalt chloride. MAIN OUTCOME MEASURE(S): Pooled FF vascular endothelial growth factor/vascular permeability factor concentrations and media vascular endothelial growth factor/vascular permeability factor accumulation at 24 and 48 hours were determined. RESULT(S): Follicular fluid vascular endothelial growth factor/vascular permeability factor concentrations were higher in advanced reproductive age women compared with younger women (3,735 +/- 2,155 versus 2,205 +/- 952 pg/mL, mean +/- SD). Accumulation of vascular endothelial growth factor/vascular permeability factor at 24 and 48 hours was 391 +/- 54 and 744 +/- 2 pg/mL in media maintained in 5% CO2 and air. Cobalt chloride induced a marked increase in vascular endothelial growth factor/vascular permeability factor (2,008 +/- 52 pg/mL at 24 hours and 3,630 +/- 519 pg/mL at 48 hours). An intermediate but significant increase in vascular endothelial growth factor/vascular permeability factor (733 +/- 35 pg/mL at 24 hours and 2,675 +/- 864 pg/mL at 48 hours) was observed with 1% O2 compared with normoxic controls. CONCLUSION(S): After hMG and hCG administration the FF from women of advanced reproductive age showed increased vascular endothelial growth factor/vascular permeability factor concentrations compared with younger women. Increased vascular endothelial growth factor/vascular permeability factor concentrations could be consistent with a hypoxic environment within follicles of older women.

Interleukin-1 beta modulates prostaglandin and progesterone production by primate luteal cells in vitro.

Increasing evidence suggests that cytokine products of the immune system may play a regulatory role in corpus luteum regulation in several species. The role of cytokines in primate luteal function, however, remains unclear. In the present study we examined the effects of interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF alpha), and interferon-gamma (IFN-gamma) on progesterone and prostaglandin (PGE2, PGF2 alpha) production by primate luteal cells in vitro. Specifically, corpora lutea were removed from normally cycling cynomolgus monkeys (n = 30 corpora lutea) during either the early (Days 3-5 after the estimated LH surge), mid (Days 8-10), or late (Days 12-14) luteal phase of the menstrual cycle. The corpora lutea were dispersed into individual cells using collagenase, DNase, and hyaluronidase. Approximately 50,000 viable luteal cells per tube were incubated in Ham's F-10 medium with increasing concentrations of IL-1 beta (0.1-10 ng/ml), TNF alpha (1-100 ng/ml), or IFN-gamma (10-1000 U/ml) in the presence and absence of hCG for 8 h at 37 degrees C. TNF alpha and IFN-gamma had no effect on progesterone PGE2, or PGF2 alpha production during any phase of the cycle at the doses tested. In contrast, IL-1 beta significantly stimulated PGF2 alpha production in a dose-dependent manner during the mid and late luteal phases (p < 0.05). Human CG alone had no effect on PGE2 or PGF2 alpha production by dispersed luteal cells in vitro but inhibited IL-1 beta-stimulated PGF2 alpha production. As expected, hCG stimulated progesterone production by primate luteal cells in vitro. Interestingly, IL-1 beta inhibited this hCG stimulation of progesterone production. In summary, these date suggest that IL-1 beta is a potentially important modulator of prostaglandin production by the primate corpus luteum. In view of this, cytokine-mediated changes in prostaglandin production by the primate corpus luteum may participate in the physiological regulation of luteal function.

Identification of gonadotrophin surge-inhibiting factor (GnSIF)/attenuin in human follicular fluid.

Evidence from several laboratories suggests that the ovaries of many species produce a non-steroidal factor called gonadotrophin surge-inhibiting or attenuating factor (GnSIF) which may regulate the response of the pituitary to gonadotrophin-releasing hormone (GnRH) and as such, may modulate the timing and/or amplitude of the luteinizing hormone (LH) surge. We have recently isolated a candidate GnSIF from porcine follicular fluid (PFF). Porcine GnSiF is a 69 kDa protein which has undetectable inhibin and follistatin immunological and biological activity. The present study was designed to purify and identify GnSIF from human follicular fluid. GnSIF activity was measured as suppression of GnRH-stimulated LH secretion from rat pituitary cells in primary culture. Human follicular fluid (approximately 500 ml) was recovered from patients undergoing in-vitro fertilization (IVF). GnSIF was purified by heparin-sepharose, Q-sepharose, S-sepharose, and hydrophobic interaction chromatography followed by isoelectric focusing. Gel electrophoresis and Western blot were used to identify human GnSIF and compare it with porcine GnSIF. Using these steps, we obtained a highly-purified preparation of GnSIF that manifests in-vitro bioactivity and chromatography characteristics similar to those observed for porcine GnSIF and that hybridizes with a porcine GnSIF antibody. Following treatment with human chorionic gonadotrophin/human menopausal gonadotrophin (HMG/HCG), human follicular fluid contained roughly 25% of the GnSIF (per mg protein) present in porcine follicular fluid. We conclude that GnSIF is present in human follicular fluid and may participate in the regulation of gonadotrophin secretion in this species.

Regulation of cumulus cell steroidogenesis by the porcine oocyte and preliminary characterization of oocyte-produced factor(s).

This study was designed to investigate whether porcine oocytes produce a factor(s) that influences cumulus and mural granulosa cell steroid production and to characterize the biochemical nature and mode of action of a such factor(s). Porcine cumulus-oocyte complexes (COC) were collected from 2-5 mm follicles and cultured either intact or after oocytectomy for 48 h. Steroid levels were then measured in the culture media. Conditioned media, obtained by culturing denuded oocytes for 48 h, were subjected to heat treatment of charcoal extraction and utilized to culture intact and oocytectomized COC. FSH-stimulated progesterone, 20 alpha-OH-progesterone, and estradiol were significantly higher in oocytectomized vs. intact COC cultures. Denuded oocytes cultured with granulosa cells significantly inhibited progesterone production compared to control. Also, media conditioned with different numbers of denuded oocytes (0 to 300) significantly inhibited progesterone production by oocytectomized COC in a manner dependent on oocyte number. Charcoal extraction, but not heat treatment, significantly removed the inhibitory effect of the conditioned media on progesterone production by oocytectomized COC. Increased progesterone production by oocytectomized COC was not accompanied by a similar increase in cAMP formation. Heptanol, a gap junction blocker, did not alter progesterone production by intact COC. In conclusion, porcine oocytes secrete a factor(s) that inhibits cumulus and mural granulosa cell steroidogenesis. This factor(s) is heat stable but extractable by charcoal. The factor(s) appears not to be transferred to somatic cells via gap junctions, and its effect is downstream of cAMP formation.

Interleukin-1 beta induces cyclooxygenase-2 in cultured human decidual cells.

PROBLEM: The purpose of this study was to examine the hypothesis that interleukin-1 beta (IL-1 beta)-elicited increases in decidual prostaglandin E2 and F2 alpha (PGE2 and PGF2 alpha) biosynthesis are due to the de novo expression of the inducible isoform of cyclooxygenase (i.e., COX-2). METHOD: Primary human decidual cell cultures were established from term placentas delivered by cesarean section. After 8 days in vitro, when the cultures secreted immunoreactive prolactin, the cells were incubated for 24 h in serum-free medium, and then challenged with IL-1 beta from 1 to 48 h. PGE2 and PGF2 alpha content in the media were measured by specific radioimmunoassays. RESULTS: IL-1 beta stimulated a time-dependent enhancement in PGE2 and PGF2 alpha production, with PGF2 alpha synthesis predominating over PGE2. IL-1 beta also induced a dose-dependent increase in the output of both arachidonic acid metabolites. When Northern blots of IL-1 beta-treated and control cells were probed with cDNAs encoding either COX-1 or COX-2 isoforms or an oligonucleotide probe encoding a portion of the human beta-actin, we detected a time- and dose-dependent increase in the steady-state levels of COX-2, but not COX-1 or beta-actin mRNA transcripts. Moreover, the expression of COX-2 mRNA in IL-1 beta-stimulated cells was superinduced by preincubation with cycloheximide, but completely abolished by actinomycin D. CONCLUSIONS: Taken together, the data suggest that COX-2 mRNA expression is largely responsible for the robust increase in PG formation seen in IL-1 beta-treated decidual cells.

Noncanalization of the fallopian tube. A case report.

Anomalous development of the fallopian tubes is an uncommon occurrence, and only a few types of structural changes have been described. A woman with a unicornuate uterus and noncanalized contralateral fallopian tube with absence of the proximal segment is described. This condition has not been reported previously.

Effect of hormonal manipulation on human fallopian tubal epithelium in vitro.

PURPOSE: Estradiol (E2) induced ciliogenesis has been demonstrated in several mammalian species. There is no consensus as to what extent, if any, this occurs in humans. The aim of this study was to determine whether hormonal manipulation could limit deciliation and/or stimulate ciliogenesis using human tubal epithelium in vitro. MATERIALS AND METHODS: Experiment 1: Tubal epithelial organ explant cultures were established from nine women undergoing hysterectomy. Several segments from each tube were fixed immediately (in vivo control), the remainder were maintained in culture for one week. One group received no hormontal treatment (in vitro control). The others were supplemented with E2 2 ng/ml (low E2) or 10 ng/ml (high E2), Clomiphene citrate 300 ng/ml + low E2 (CC), and progesterone 300 ng/ml + low E2 (P4). All specimens were examined by scanning electron microscopy and the percent ciliated area determined. Experiment 2: Tubal epithelial cultures were established from an additional 11 patients. After 1 week in culture, half of the explants from each patient were supplemented with E2 15 ng/ml vs no E2 and maintained for another week. The specimens were examined as above, as well as with transmission electron microscopy (TEM). RESULTS: Experiment 1: After 1 week in culture the mean percent ciliated areas in the in vitro control (24.8 +/- 17.9), low E2 (24.7 +/- 17.0), P4 (20.7 +/- 10.7), and CC (25.8 +/- 17.2) were approximately half the in vivo control (45.7 +/- 10.4), P = 0.005. The high E2 group (39.7 +/- 19.7) was significantly higher than the in vitro control, P = 0.008, but was not different from the in vivo control. Experiment 2: Both groups demonstrated a great reduction in ciliated area and were not significantly different from each other, E2 (8.3 +/- 10.8) and control (3.0 +/- 3.0). TEM failed to demonstrate ciliogenic precursors in either group. CONCLUSIONS: High E2 was capable of preventing initial epithelial deciliation in vitro. Once deciliation started however, high E2 was unable to limit the process or induce ciliogenesis.

Juxtaposition of contralateral ovary and fallopian tube to allow pregnancy in unicornuate uterine anomaly.

Infertile patients who have undergone unilateral oophorectomies or salpingectomies or those with uterine anomalies may require juxtaposition of a remaining fallopian tube and contralateral ovary. A 28-year-old woman with previous left oophorectomy for endometriosis was found to have a left unicornuate uterus during subsequent infertility evaluation. Laparotomy and resection of a right rudimentary horn and juxtaposition of the left fallopian tube and the right ovary was performed. A large endometrioma was also resected from the right ovary. The patient conceived during the second month postoperatively. A simple procedure for juxtaposing a contralateral fallopian tube and ovary is described. Careful evaluation of the pelvic organs should be performed before removal of an ovary in a young woman.

Recombinant human follicle-stimulating hormone stimulates multiple follicular growth, but minimal estrogen production in gonadotropin-releasing hormone antagonist-treated monkeys: examining the role of luteinizing hormone in follicular development and steroidogenesis.

The two-cell theory predicts that follicular steroidogenesis requires the coordinate actions of both FSH and LH; however, the role of LH in follicular growth is less clear. The present study was designed to investigate the relative importance of LH and FSH in follicular growth and steroidogenesis. Cynomolgus monkeys were treated with a GnRH antagonist (antide; 3 mg/kg.day) for 20 days beginning in the midluteal phase of the menstrual cycle. After 10 days of antide administration, monkeys were injected with recombinant human FSH (rhFSH; 10 IU; n = 3), human menopausal gonadotropin (hMG; 10 IU; n = 3), or FSH plus 0.5 IU LH (n = 3) twice daily for 10 days. rhFSH stimulated multiple follicular development; however, peak serum estradiol levels were only 943 +/- 195 pmol/L. In contrast, monkeys treated with the same dose of hMG had significantly higher (P < 0.05) peak estradiol levels (6013 +/- 1322 pmol/L). The addition of 0.5 IU LH to the rhFSH treatment resulted in serum estradiol levels similar to those in monkeys treated with rhFSH only. Importantly, no differences in follicle number or size were evident among these treatment groups. Follicular fluid estradiol levels were consistent with serum levels (rhFSH, 187 +/- 11 nmol/L; hMG, 1531 +/- 173 nmol/L). Even larger proportional differences in follicular fluid androstenedione (rhFSH 13.6 +/- 1.4 nmol/L; hMG, 307 +/- 97.7 nmol/L) levels were found. The results in this LH-deficient primate model suggest that FSH alone is capable of stimulating ovarian follicular growth; however, the resulting follicles manifest minimal estradiol production, probably due to deficiencies in the LH-induced precursors to estradiol.

Novel human endometrial cell line promotes blastocyst development.

OBJECTIVE: To test and contrast the embryotrophic potential of an established human endometrial cell line to that of two other epithelial cell types: human oviduct and African monkey kidney (Vero) cells. DESIGN: Mouse IVF was performed. Subsequent development of embryos cocultured with our endometrial cell line was contrasted to that seen with oviductal and Vero cell coculture systems. Percent blastocyst transformation, expansion, and hatching were compared. SETTING: University-based research laboratory associated with clinical IVF program. RESULTS: All three epithelial cell monolayers tested significantly improved the rate of blastocyst transformation of in vitro fertilized murine oocytes. The overall percent blastocysts obtained was highest with endometrial cells (69%), followed by oviductal cells (52%), Vero cells (45%), and the medium-alone controls (29%). Only 13% of oviduct cocultured embryos were able to reach the hatched blastocyst stage compared with a 30% hatching rate with endometrial cells and a 21% hatching rate with Vero cells. Only 3% of control embryos hatched in vitro. CONCLUSION: We have described a novel continuous endometrial cell line with excellent embryotrophic potential. This cell line is technically easy to use and is of human origin. As a coculture system it appears to be superior to both oviductal and Vero cells in correcting for defects in culture environment during in vitro development up to the blastocyst stage.

Pregnancy can be established in superovulated adult rats treated with progesterone and an aromatase inhibitor.

An adult superovulated rat model has been developed and is characterized by high ovulation rates, early morphological degeneration of embryos, complete embryo loss within 48 hours of conception and elevated peripheral estradiol(E2)/progesterone(P4) ratios. In this study, three trials were conducted using the superovulated adult rat model. First, control naturally cycling rats were compared with superovulated rats supplemented with 1 mg P4 on days 0-3 of pregnancy. A sperm positive vaginal smear is designated as day 0 of pregnancy. The P4 treated rats demonstrated improved embryo retrieval on day 1 of pregnancy, continued embryo recovery with a decrease in normal morphologic characteristics of integrity on day 2, with nearly total embryo loss by day 3. On each day, P4 levels were elevated 2-3 times over control. The second trial compared 3 groups of rats, 1) naturally cycling, 2) superovulated unsupplemented and 3) superovulated rats supplemented with 1 mg P4/rat/day and the aromatase inhibitor, 4-hydroxyandrostenedione (4-OHA), 12.5 mg/rat/day. The superovulated unsupplemented rats had no embryo recovery after day 2 of pregnancy, while the P4 and 4-OHA treated rats showed a variable ability to maintain normally developing embryos through day 4 of pregnancy. E2 levels were elevated in both superovulated groups on days 1-4 of pregnancy as were P4 levels on days 2-4. The E2/P4 ratio was significantly lowered only on day 1 of pregnancy in the P4 and 4-OHA treated group. The third trial demonstrated implantation in 50% of the superovulated rats supplemented with P4 and 4-OHA. In conclusion, implantation in the superovulated adult rats can occur with P4 and 4-OHA supplementation, however, this biologic phenomenon could not be explained by obvious changes in peripheral E2 and P4 levels.

The effect of preovulatory peritoneal fluid from cases of endometriosis on murine in vitro fertilization, embryo development, oviduct transport, and implantation.

OBJECTIVE: The null hypothesis of our study is that the success of in vitro and in vivo murine fertilization and embryo development is not decreased by gamete exposure to peritoneal fluid from superovulated patients with endometriosis. STUDY DESIGN: A murine in vitro fertilization model was used to test the effects of endometriosis versus nonendometriosis peritoneal fluid at concentrations of 1%, 5%, and 10% versus an unsupplemented control. Fertilization and blastocyst formation were compared by analysis of variance. In a second experiment superovulated mice were given intraperitoneal injections of endometriosis or nonendometriosis fluid or saline solution 8 hours after human chorionic gonadotropin and then mated. Some mice were killed 3 days after coitus to assess embryo number, cleavage stage, and uterine versus tubal position by means of analysis of variance and covariance with repeated measures. Others were killed 12 days after coitus with the mean number of implantations per animal between groups compared by Student's t test. RESULTS: In vitro fertilization rates decreased as peritoneal fluid concentration increased in both the endometriosis (65%, 43%, 33%) and nonendometriosis (65%, 52%, 35%) groups at 1%, 5%, and 10% peritoneal fluid concentration, respectively. Mice receiving intraperitoneal endometriosis or nonendometriosis fluid or saline solution injections showed no differences in embryo number, cleavage, uterine versus tubal position, or mean implantation number. CONCLUSION: Peritoneal fluid from superovulated patients had no differentially negative effect when compared with the effect of nonendometriosis peritoneal fluid on murine in vitro or in vivo fertilization and embryo development, tubal embryo transport, or implantation.

Improvement of in vitro fertilization and early embryo development in mice by coculture with human fallopian tube epithelium.

Coculturing one- and two-cell embryos with various cell lines has been shown to overcome species-specific developmental blocks and to improve blastocyst transformation rates. The objective of this study was to assess whether human fallopian tube epithelium organ explants influence in vitro fertilization and subsequent early embryo development in a murine model. Fertilization, blastocyst transformation, and blastocyst expansion and hatching rates were significantly higher in the coculture group when compared with rates for culture in standard media or media conditioned by human tubal explant cultures. The results from conditioned and unconditioned media were not significantly different.

Effect of baseline ovarian cysts on in vitro fertilization and gamete intrafallopian transfer cycles.

The presence of ovarian cysts may compromise the success of in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT). We prospectively studied 212 consecutive ovulation induction cycles in 120 patients for IVF and/or GIFT. A baseline cyst was defined as any intraovarian cystic structure greater than or equal to 12 mm noted on ultrasonography before superovulation. Cycle outcomes were compared between patients with cysts (n = 62) versus those with no cysts (n = 150). There were no differences in follicular or luteal phase lengths or amount of human menopausal gonadotropins used. Peak estradiol (E2) levels were significantly lower and cancellation rates significantly higher in the cyst group. For noncanceled cycles, there were no significant differences in peak E2 levels, the mean number of follicles greater than or equal to 12 mm, mature oocytes retrieved, or ova transferred for GIFT or embryos for IVF. The pregnancy rates overall and for noncanceled cycles were not significantly different.