Novel extra cellular-like matrices to improve human ovarian grafting.

PURPOSE: To investigate if human ovarian grafting with pure virgin human recombinant collagen type-1 from bioengineered plant lines (CollPlant™) or small intestine submucosa (SIS) yields better implantation results for human ovarian tissue and which method benefits more when combined with the host melatonin treatment and graft incubation with biological glue + vitamin E + vascular endothelial growth factor-A. METHODS: Human ovarian tissue wrapped in CollPlant or SIS was transplanted into immunodeficient mice with/without host/graft treatment. The tissue was assessed by follicle counts (including atretic), for apoptosis evaluation by terminal deoxynucleotidyl transferase assay and for immunohistochemical evaluation of neovascularization by platelet endothelial cell adhesion molecule (PECAM) expression, and for identification of proliferating granulosa cells by Ki67 expression. RESULTS: Human ovarian tissue transplanted with CollPlant or SIS fused with the surrounding tissue and promoted neovascularization. In general, implantation with CollPlant even without additives promoted better results than with SIS: significantly higher number of recovered follicles, significantly fewer atretic follicles, and significantly more granulosa cell proliferation. Moreover, results with CollPlant alone seemed to be at least as good as those after host and graft treatments. CONCLUSIONS: CollPlant is a biomaterial without any potential risks, and grafting ovarian tissue with CollPlant is easy and the procedure may be easily modified, with limited or no foreseeable risks, for auto-transplantation in cancer survivors. Further studies are needed using other novel methods capable of enhancing neovascularization and reducing apoptosis and follicle atresia.

Use of Simvastatin, Fibrin Clots, and Their Combination to Improve Human Ovarian Tissue Grafting for Fertility Restoration After Anti-Cancer Therapy.

Anticancer treatments, particularly chemotherapy, induce ovarian damage and loss of ovarian follicles. There are limited options for fertility restoration, one of which is pre-chemotherapy cryopreservation of ovarian tissue. Transplantation of frozen-thawed human ovarian tissue from cancer survivors has resulted in live-births. There is extensive follicular loss immediately after grafting, probably due to too slow graft revascularization. To avoid this problem, it is important to develop methods to improve ovarian tissue neovascularization. The study's purpose was to investigate if treatment of murine hosts with simvastatin or/and embedding human ovarian tissue within fibrin clots can improve human ovarian tissue grafting (simvastatin and fibrin clots promote vascularization). There was a significantly higher number of follicles in group A (ungrafted control) than in group B (untreated tissue). Group C (simvastatin-treated hosts) had the highest levels of follicle atresia. Group C had significantly more proliferating follicles (Ki67-stained) than groups B and E (simvastatin-treated hosts and tissue embedded within fibrin clots), group D (tissue embedded within fibrin clots) had significantly more proliferating follicles (Ki67-stained) than group B. On immunofluorescence study, only groups D and E showed vascular structures that expressed both human and murine markers (mouse-specific platelet endothelial cell adhesion molecule, PECAM, and human-specific von Willebrand factor, vWF). Peripheral human vWF expression was significantly higher in group E than group B. Diffuse human vWF expression was significantly higher in groups A and E than groups B and C. When grafts were not embedded in fibrin, there was a significant loss of human vWF expression compared to groups A and E. This protocol may be tested to improve ovarian implantation in cancer survivors.

The influence of in vivo exposure to nonylphenol ethoxylate 10 (NP-10) on the ovarian reserve in a mouse model.

AIM: To determine the effect of nonylphenol-ethoxylate-10 (NP-10) on the ovarian reserve in a mouse model. DESIGN: Female mice were maintained on purified water or exposed to NP-10 from 3-7-weeks of age. At 7-weeks they were stimulated, mated and the zygotes were cultured in-vitro. Three and 7-weeks old mice were untreated controls. Identical groups were sacrificed without stimulation. Ovaries were analysed for follicular composition. Respiratory-chain (RC) activity and reactive-oxygen-species (ROS) production were measured in brains and livers. RESULTS: Seven-weeks-old mice produced fewer oocytes/embryos than 3-week-old mice. At 7-weeks, mice exposed to NP-10 produced more oocytes/embryos the controls. Their ovaries contained more primordial/primary follicles, with a lower rate of proliferation and fewer antral follicles. There were no differences in follicular apoptosis, RC-activity or ROS production. CONCLUSIONS: In this model, exposure to NP-10 inhibited the spontaneous follicular recruitment, the first report of successful inhibition of physiologic ovarian aging, to the best of our knowledge.

Endometrial thickness of less than 7.5 mm is associated with obstetric complications in fresh IVF cycles: a retrospective cohort study.

RESEARCH QUESTION: Does endometrial thickness affect the occurrence of obstetric complications in fresh IVF cycles? DESIGN: We conducted a retrospective cohort study that included all singleton deliveries resulting from fresh embryo transfers in a single centre between 2008 and 2014. Obstetric complications, i.e. preeclampsia, placental abruption, placenta previa, small for gestational age and preterm delivery, in singleton live births were compared among patients with an endometrial thickness of less than 7.5 mm and 7.5 mm or over on day of HCG triggering. We adjusted for confounders, including maternal age, body mass index, smoking, peak oestradiol, parity, chronic hypertension, pre-gestational diabetes, gestational diabetes, vanishing twin, inherited or acquired thrombophilia, and past pregnancy complications. RESULTS: A total of 5546 fresh embryo transfer cycles were carried out during the study period, of which 864 singleton deliveries met inclusion criteria. After adjusting for potential confounders, an endometrial thickness of less than 7.5 mm was found to be associated with increased risk for adverse obstetric outcome (adjusted OR 1.53; 95% CI 1.03 to 2.42; P = 0.04) even after excluding patients with prior pregnancy complications (adjusted OR 2.2; 95% CI 1.05 to 4.59; P = 0.035). CONCLUSIONS: Our results demonstrated that a thin endometrial lining was associated with obstetric complications that might be related to poor placentation. These findings should be validated in large prospective cohort studies.

Use of Aromatase Inhibitors in IVF for Fertility Preservation of Non-Breast Cancer Patients: A Case Series.

BACKGROUND: Controlled ovarian hyperstimulation (COH) followed by oocyte retrieval is a leading option for fertility preservation before chemotherapy, yet this procedure causes excessive serum levels of estradiol (E2), which are often detrimental for cancer patients. Aromatase inhibitors are often used in breast cancer patients during COH to prevent elevated levels of E2. OBJECTIVES: To describe our experience with COH for oocyte cryopreservation in non-breast cancer patients using aromatase inhibitors. METHODS: Of the five patients treated, two had an aggressive abdominal desmoid tumor, one had endometrial carcinoma, one had uterine sarcoma, and one patient had a brain oligodendroglioma. In all cases the treating oncologist suggested an association between estrogen and possible tumor progression. All patients were treated with a standard in vitro fertilization antagonist protocol combined with aromatase inhibitors, similar to the protocol used for breast cancer patients. RESULTS: The average duration of treatment was 10.5 days, mean peak E2 was 2348 pmol/L, mean number of oocytes aspirated was 17.3, and a mean of 14.6 embryos/oocytes were cryopreserved. CONCLUSIONS: COH with aromatase inhibitors is apparently effective in non-breast cancer patients and spares exposure to high E2 levels.

Female fertility preservation: past, present and future.

Anti-cancer therapy, particularly chemotherapy, damages ovarian follicles and promotes ovarian failure. The only pharmacological means for protecting the ovaries from chemotherapy-induced injury is gonadotrophin-releasing hormone agonist, but its efficiency remains controversial; ovarian transposition is used to shield the ovary from radiation when indicated. Until the late 1990s, the only option for fertility preservation and restoration in women with cancer was embryo cryopreservation. The development of other assisted reproductive technologies such as mature oocyte cryopreservation and in vitro maturation of oocytes has contributed to fertility preservation. Treatment regimens to obtain mature oocytes/embryos have been modified to overcome various limitations of conventional ovarian stimulation protocols. In the last decades, several centres have begun cryopreserving ovarian samples containing primordial follicles from young patients before anti-cancer therapy. The first live birth following implantation of cryopreserved-thawed ovarian tissue was reported in 2004; since then, the number has risen to more than 130. Nowadays, ovarian tissue cryopreservation can be combined with in vitro maturation and vitrification of oocytes. The use of cryopreserved oocytes eliminates the risk posed by ovarian implantation of reseeding the cancer. Novel methods for enhancing follicular survival after implantation are presently being studied. In addition, researchers are currently investigating agents for ovarian protection. It is expected that the risk of reimplantation of malignant cells with ovarian grafts will be overcome with the putative development of an artificial ovary and an efficient follicle class- and species-dependent in vitro system for culturing primordial follicles.

[LAPAROSCOPICALLY-ASSISTED ULTRASOUND-GUIDED PERCUTANEOUS TRANSABDOMINAL OOCYTE COLLECTION: FERTILITY PRESERVATION IN A 17-YEARS-OLD GIRL WITH VAGINAL EWING SARCOMA].

INTRODUCTION: Options for preserving fertility in children and adolescents with cancer depend on patient age, the available time frame, and the treatment regimen. Ovarian stimulation with mature oocyte preservation is often the optimal method in post-menarcheal adolescents. We describe a case of a 17-year-old girl with vaginal soft-tissue Ewing sarcoma in whom transvaginal oocyte collection for fertility preservation was ruled out by the large tumor. To overcome the limitations of the transabdominal approach, we applied a novel method of laparoscopically-assisted ultrasound-guided percutaneous transabdominal oocyte collection. In this manner, we were able to both perform oophorectomy and obtain superficial and deep ovarian follicles for cryopreservation.

Fertility Treatment Outcomes After Follicle Tracking With Standard 2-Dimensional Sonography Versus 3-Dimensional Sonography-Based Automated Volume Count: Prospective Study.

OBJECTIVES: The use of sonography-based Automated Volume Count (SonoAVC; GE Healthcare, Kretz, Zipf, Austria) leads to substantially lower intraobserver and interobserver variability with a considerable advantage in time gain for both the physician and patient. It offers the possibility of continuous training and standardization of follicular monitoring. Manual and automated follicular measurements during in vitro fertilization (IVF) are reported to be comparable during gonadotropin-releasing hormone (GnRH) agonist treatment. The aim of our study was to evaluate the effect of follicle tracking with 3-dimensional (3D) SonoAVC on treatment outcomes in GnRH antagonist IVF cycles. METHODS: A prospective trial included 54 women undergoing their first to fourth GnRH antagonist IVF cycles. Follicle tracking from the initiation of ovarian stimulation until the day of oocyte retrieval and timing of oocyte retrieval was done either by conventional 2-dimensional (2D) sonography or 3D SonoAVC (open-labeled parallel assignment). In both groups, recombinant human chorionic gonadotropin was injected when there were at least 3 leading follicles measuring 17 mm. The primary outcome was the oocyte maturation rate, and secondary outcomes were the fertilization rate and clinical pregnancy rate. RESULTS: The number of retrieved oocytes, number and rate of mature oocytes, fertilization rate, and clinical pregnancy rate were similar for 2D sonography and 3D SonoAVC. On a multivariate regression analysis, the use of 3D sonography was not a significant independent predictor of mature oocytes or clinical pregnancy rates. CONCLUSIONS: Follicle tracking with 3D sonographic follicular volume measurements does not achieve better fertility outcomes than standard 2D sonography.

First-trimester inter- and intrafetal size discrepancies in bichorionic twins conceived by in vitro fertilization: can it predict pregnancy outcome?

OBJECTIVE: To evaluate the association between first-trimester inter- and intrafetal size discrepancies and pregnancy outcome, among bichorionic-biamniotic twins conceived by IVF. DESIGN: A retrospective study design was used. SETTING: Tertiary university-affiliated medical center. PATIENT(S): All women with a viable first-trimester bichorionic-biamniotic twin gestation, who conceived after IVF in 2007-2015. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The association between fetal size differences and pregnancy outcome was analyzed. Intrafetal size discordance was defined as a difference between the actual gestational age calculated by ovum pickup (OPU) date and the evaluated gestational age by crown-rump length (CRL), for each twin. Intertwin size discrepancy was defined as a difference in CRL between the twins. The primary outcome was the number of live-born fetuses; the secondary outcome measures were gestational age at birth, birth weight percentile, and birth weight discordancy. RESULT(S): A total of 277 women met the study criteria and were divided into three groups by outcome: 218 (78.7%) live-born twins, 41 (14.8%) live-born singleton, and 19 (6.5%) non-live-born pregnancy. Among the smaller than expected twin, the association of CRL-OPU differences with the primary outcome was significant for twin live-born delivery (-1.43 day), singleton live-born delivery (-4.12 days), and non-live-born pregnancy (-6.72 days). For the relatively larger twin, the association was significant for non-live-born pregnancy (-4.33 days) compared with any live-born delivery, either singleton (-0.95 days) or twin (-0.21 days). CONCLUSION(S): Among IVF conceived twin gestations, a CRL-OPU gap was associated with an increased risk of a negative pregnancy outcome.

[FERTILITY PRESERVATION IN YOUNG CANCER PATIENTS - CAN WE OPTIMIZE THE PATH?]

INTRODUCTION: Advances in cancer therapy have improved the long-term survival of cancer patients. Concerns about fertility represent a major issue for young cancer patients. The emergent discipline of oncofertility, an intersection between oncology and fertility, is a new concept that describes an integrated network of clinical resources that focus on fertility preservation from both clinical and research perspectives. Patients and methods: In this article we describe our designated multidisciplinary program for fertility preservation in pediatric and young adult populations. The program is also designed to serve as a prospective platform for the evaluation of reproductive outcomes in this patient cohort. RESULTS: We have observed considerably higher referral rates following launching the program and earlier referral of chemonaïve patients that concedes maximal fertility preservation. Two hundred and thirty five patients were referred to the program over a period of 3 years. CONCLUSIONS: Our program demonstrates that multidisciplinary programs that encompass relevant specialists, skilled laboratory resources and a facilitated path that drives the process in the shortest time, maximizes the yield.

Attempts to improve human ovarian transplantation outcomes of needle-immersed vitrification and slow-freezing by host and graft treatments.

PURPOSE: To investigate if needle-immersed vitrification or slow-freezing yields better implantation results for human ovarian tissue and which method benefits more when combined with the "improvement protocol" of host melatonin treatment and graft incubation with biological glue + vitamin E + vascular endothelial growth factor-A. METHODS: Human ovarian tissue was preserved by needle-immersed vitrification or slow-freezing and transplanted into immunodeficient mice, either untreated (groups A and C, respectively) or treated with the improvement protocol (groups B and D, respectively). Grafted and ungrafted slices were evaluated by follicle counts, apoptosis assay and immunohistochemistry for Ki67 and platelet endothelial cell adhesion molecule (PECAM). RESULTS: Follicle number in the recovered grafts was limited. The number of atretic follicles was significantly higher after vitrification with/without the improvement protocol and slow-freezing than that after slow-freezing + the improvement protocol. Stroma cell apoptosis was the lowest in the group D. PECAM staining showed a peripheral and diffuse pattern in the group D (mostly normal follicular morphology) and a diffuse pattern in all other groups (few follicles, mostly atretic), with significantly higher diffuse levels in the vitrification groups. Ki67 staining was identified in all normal follicles. Follicles did not survive transplantation in the vitrification groups. CONCLUSIONS: Ovarian sample preparation with slow-freezing + the improvement protocol appears to yield better implantation outcomes than needle-immersed vitrification with/without the improvement protocol. The real quality of frozen tissue can be assessed only after grafting and not after thawing/warming.

Extracellular-like matrices and leukaemia inhibitory factor for in vitro culture of human primordial follicles.

The possibility of maturing human primordial follicles in vitro would assist fertility restoration without the danger of reseeding malignancies. Leukaemia inhibitory factor (LIF) and certain culture matrices may promote human follicular growth. The present study compared human primordial follicular growth on novel culture matrices, namely human recombinant vitronectin (hrVit), small intestine submucosa (SIS), alginate scaffolds and human recombinant virgin collagen bioengineered in tobacco plant lines (CollPlant). The frozen-thawed ovarian samples that were used had been obtained from girls or young women undergoing fertility preservation. In the first part of the study, 20 samples were cultured for 6 days on hrVit or SIS with basic culture medium alone or supplemented with one of two concentrations of LIF (10ngmL-1 and 100ngmL-1), with and without LIF-neutralising antibody. In the second part of the study, 15 samples were cultured for 6 days on alginate scaffolds or CollPlant matrices with basic culture medium. Follicular development was assessed by follicular counts and classification, Ki67 immunohistochemistry and 17ß-oestradiol and anti-Müllerian hormone measurements in spent media samples. Primordial follicular growth was not enhanced by LIF. Despite some significant differences among the four matrices, none appeared to have a clear advantage, apart from significantly more Ki67-stained follicles on alginate and CollPlant matrices. Further studies of other culture matrices and medium supplements are needed to obtain an optimal system.

Prediction of fetal loss by first-trimester crown-rump length in IVF pregnancies.

OBJECTIVE: To evaluate the association between small crown-rump length (CRL) and fetal loss ≤22 weeks in IVF pregnancies. METHODS: A retrospective analysis of prospectively collected data at a university-affiliated medical center. All singleton IVF pregnancies within a 5-year period, with a live embryo on first-trimester ultrasound and verified pregnancy outcome were included. Rates of fetal loss ≤22 weeks were compared between pregnancies with a CRL ≤tenth percentile and above the tenth percentile of our population. RESULTS: Overall, 397 pregnancies met inclusion criteria. Ninety-five percent of CRL measurements were performed at 40-80 gestational days. All live-embryo's CRL measurements, from 40 to 80 mm, were plotted against expected gestational age (in 5-day clusters), with calculation of the tenth percentile for every gestational age. Total of 64 pregnancies had CRL ≤tenth percentile for gestational age. The rate of fetal loss in this group was significantly higher than in pregnancies with CRL >tenth percentile (17.2 vs. 6.6%, p = 0.005, OR = 2.93, 95% CI 1.2-6.7). In both groups, the majority of fetal losses occurred ≤10 weeks of gestation. CONCLUSION: In IVF pregnancies with a live embryo, a small CRL at 40-80 days' gestation may predict fetal loss. Repeated ultrasound should be considered after 1-2 weeks.

Aspiration of immature oocytes during cesarean section for fertility preservation.

BACKGROUND: In vitro maturation (IVM) of immature oocytes is an important technology for selected clinical indications. We previously described a pregnant woman with a history of renal transplantation who underwent oocyte aspiration during cesarean section (CS) for fertility preservation and future surrogacy. CASE: A 27-year-old pregnant woman was diagnosed with neck rhabdomyosarcoma at 37 weeks' gestation. CS was performed with direct aspiration of small follicles from one ovary and oophorectomy of the other. Twenty-one identified oocyte-cumulus complexes were cultured, and 12 mature oocytes and 14 ovarian cortex strips were cryopreserved. CONCLUSION: Aspirating competent oocytes during CS may serve as an additional means of fertility preservation in pregnant women. The procedure may also be offered to patients with an IVF pregnancy who are scheduled for elective CS.

Short-term exposure of human ovarian follicles to cyclophosphamide metabolites seems to promote follicular activation in vitro.

How chemotherapy affects dormant ovarian primordial follicles is unclear. The 'burnout' theory, studied only in mice, suggests cyclophosphamide enhances primordial follicle activation. Using 4-hydroperoxycyclophosphamide (4hc) and phosphoramide mustard (PM), this study assessed how the active cyclophosphamide metabolites 4-hydroxycyclophosphamide (4-OHC) and PM, affect human primordial follicles. Frozen-thawed human ovarian samples were sliced and cultured with basic culture medium (cultured controls) or with 4hc/PM (3 µmol/l/10 µmol/l) (treated samples) for 24-48 h. Follicular counts and classification, Ki67 and anti-Müllerian hormone (AMH) immunohistochemistry and an apoptosis assay were used for evaluation, and 17ß-oestradiol and AMH were measured in spent media samples. Generally, there was primordial follicle decrease and elevated developing follicle rates in treated samples compared with cultured (P = 0.04 to P < 0.0005) and uncultured controls (P < 0.05 to P < 0.0001). No traces of apoptosis were found. There were almost twicethe levels of AMH and 17ß-oestradiol in treated compared with untreated samples (AMH with 4hc 3 µmol/l; P = 0.04). All follicles stained positively for AMHincluded treated samples. Ki67 positive staining was noted in all samples. Cyclophosphamide metabolites seem to enhance human primordial follicle activation to developing follicles, in vitro. Study findings support the 'burnout' theory as the mechanism of chemotherapy-induced ovarian toxicity.

The effect of in vitro fertilization on coagulation parameters as measured by thromboelastogram.

OBJECTIVES: In vitro fertilization (IVF) induced elevated estrogen levels are associated with a hypercoagulable state. Thromboelastogram (TEG) is a point of care whole blood hemostasis analyzer which measures functionality of clotting parameters. Our study's objective was to examine the influence of the early and late follicular phase of an IVF simulation cycle on coagulation parameters as measured by TEG and to evaluate the influence of age on coagulation parameters. STUDY DESIGN: In a single center, prospective, observational trial, 46 women undergoing IVF therapy were studied. All women received a standardized IVF treatment protocol. Venous blood was drawn on the first day of the stimulation cycle and on the day of hCG injection and assessed by TEG. Parameters assessed by were R (represent clotting time), K and Angle (reflect clot strength and development), MA (maximum platelet-fibrin clot strength), CI (represents overall coagulability), and LY30 (represents lysis). RESULTS: Data from 46 women were analyzed. A statistically significant difference was found in all TEG parameters between early and late follicular phase, indicating a hypercoagulable state. R (p<0.0001), K (p=0.008), angle (p=0.008), MA (p=0.004), CI (p<0.001), LY30 (p=0.59). Age was a significant independent predictor for R at the early follicular phase (p=0.042). Both age and estrogen levels were found to be independent predictors for CI at late follicular phase. Age (p=0.011), estrogen (p=0.019). CONCLUSIONS: There was a significant difference in all coagulation parameters between early and late follicular phase, indicating a hypercoagulable state.

Long-Term Follow-Up of Chemotherapy-Induced Ovarian Failure in Young Breast Cancer Patients: The Role of Vascular Toxicity.

BACKGROUND: We previously reported that chemotherapy-induced ovarian toxicity may result from acute vascular insult, demonstrated by decreased ovarian blood flow and diminished post-treatment anti-Müllerian hormone (AMH) levels. In the present study, we report the continuous prospective evaluation of ovarian function in that cohort. METHODS: Patients (aged <43 years) with localized breast cancer were evaluated by transvaginal ultrasound prior to initiation of chemotherapy, immediately at treatment completion, and at 6 and 12 months after treatment cessation. Doppler flow velocity indices of the ovarian vasculature (resistance index [RI], pulsatility index [PI]) were visualized. Hormone markers of ovarian reserve were assessed at the same time points. RESULTS: Twenty patients were enrolled in the study. Median age was 34 ± 5.24 years. Ovarian blood flow was significantly reduced immediately following chemotherapy (both RI and PI; p = .01). These parameters were partially recovered at later points of assessment (6 and 12 months after treatment); patients aged <35 years significantly regained ovarian blood flow compared with patients aged >35 years (p < .05). AMH dropped dramatically in all patients following treatment (p < .001) and recovered in only 10 patients. Hormone markers of ovarian reserve shortly after chemotherapy depicted a postmenopausal profile for most patients, accompanied by related symptoms. Follicle-stimulating hormone (FSH) levels recovered in 14 of 20 patients and significantly returned to the premenopausal range in patients aged <35 years (p = .04); 10 of 20 resumed menses at 12 months. The pattern of vascular impairment was lessened in patients treated with a trastuzumab-based protocol, although results did not reach statistical significance (p = .068). CONCLUSION: Continuous prospective evaluation of ovarian vasculature and function in a cohort of young patients during and after chemotherapy indicated that ovarian toxicity may derive from acute vascular insult. Age may affect whether patients regain ovarian function, whereas recovery of blood flow and premenopausal FSH levels at later assessment was notable in patients aged <35 years. IMPLICATIONS FOR PRACTICE: This study explored the role of vascular toxicity in mediating ovarian impairment and recovery following chemotherapy. Continuous prospective evaluation of ovarian vasculature and function in a cohort of young patients during and after chemotherapy indicated that ovarian toxicity may derive from acute vascular insult. Future studies are warranted to further characterize patterns of vascular toxicity of various chemotherapies in clinical practice and to assess the role of chemotherapy-induced vascular toxicity for specific end organs such as the ovary with systemic vascular effect. Elucidating the cause of impairment may facilitate development of measures to minimize vascular toxicity and consequences of acute vascular insult.

Prolonging oocyte in vitro culture and handling time does not compensate for a shorter interval from human chorionic gonadotropin administration to oocyte pickup.

OBJECTIVE: To assess the impact of oocyte aspiration, denudation, and sperm injection timing in relation to oocyte hCG exposure time on intracytoplasmic sperm injection (ICSI) outcome. DESIGN: Cohort study. SETTING: Tertiary medical center. PATIENT(S): A total of 614 consecutive ICSI cycles were performed in 421 patients aged <38 years with at least three aspirated oocytes and no more than three previous treatments. INTERVENTION(S): Gonadotropin-releasing hormone agonist or GnRH antagonist suppression; oocyte pickup (OPU)-hCG interval more/less than 36 hours; OPU-denudation interval more/less than 2 hours; denudation-ICSI interval more/less than 1 hour. MAIN OUTCOME MEASURE(S): Fertilization, embryo transfer, and pregnancy rates. RESULT(S): Late OPU was associated with more available embryos than early OPU and significantly higher rates of fertilization (66.0% ± 22.8% vs. 61.8% ± 24.3%), ET (99.5% vs. 96.2%), and pregnancy (47.2% vs. 35.4%). This advantage was more pronounced in GnRH agonist cycles. The length of incubation before or after denudation had no effect, regardless of OPU timing. On logistic stepwise regression, OPU timing was the only significant independent predictor of pregnancy (odds ratio 1.6, 95% confidence interval 1.17-2.29). CONCLUSION(S): The timing of OPU has a predominant effect on ICSI success, especially in GnRH agonist cycles. Delaying oocyte denudation or sperm injection does not compensate for insufficient postpriming exposure to the follicular environment.

Progesterone-to-follicle index is better correlated with in vitro fertilization cycle outcome than blood progesterone level.

OBJECTIVE: To investigate the impact of late follicular phase progesterone (P) elevation in relation to ovarian response on cycle outcome. DESIGN: Cohort study. The progesterone-to-follicle index (PFI) was calculated by dividing the blood P by the number of follicles ≥14 mm. The clinical pregnancy rate was calculated against the range of PFI values and blood P levels. SETTING: In vitro fertilization unit. PATIENT(S): A heterogenous population undergoing IVF with pituitary suppression and gonadotropin stimulation resulting in 3-15 follicles ≥14 mm and blood P≤10 nmol/L on hCG day and resulting in fresh embryo transfer. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Association of blood P and PFI with clinical pregnancy rate. RESULT(S): Data were retrieved for 8,649 IVF cycles in normal responders. The (reverse) odd ratios for pregnancy were 1.112 (95% confidence interval [CI], 1.077-1.165) for blood P and 4.104 (95% CI, 3.188-5.284) for the PFI. Elevated P levels were associated with a lower pregnancy rate only when they reached the >93rd percentile. The PFI was inversely and linearly related to the pregnancy rate for the whole range of values. CONCLUSION(S): A late increase in P level is detrimental if it is a consequence of increased P production per follicle (high PFI) but not if it is a consequence of additional follicular recruitment. The PFI enables clinicians to differentiate these conditions.