Alessio Paffoni

Consistent and predictable delivery rates after oocyte vitrification: an observational longitudinal cohort multicentric study

BACKGROUND An efficient method for cryopreservation of human oocytes may offer solutions to legal and ethical problems in routine infertility programs and may also be used for fertility preservation for medical and social reasons. METHODS We conducted an observational longitudinal cohort multicentric study to investigate the efficacy and reproducibility of oocyte cryopreservation outcomes in IVF/ICSI cycles. Moreover, the effects of patient and cycle characteristics on the delivery rate (DR) were analyzed. RESULTS In 486 cycles performed in 450 couples, 2721 oocytes were warmed and 2304 of them survived cryopreservation (84.7%). Of the 2182 oocytes subjected to ICSI, the rates of fertilization and development to top-quality embryos were 75.2 and 48.1%, respectively. A total of 128 deliveries were obtained (26.3% per cycle and 29.4% per transfer) for 450 patients (28.4%) and 147 babies were live born from 929 embryos transferred (15.8%). The forward logistic regression analysis on a per patient basis showed that female age [odds ratio (OR): 0.93, 95% confidence interval (CI): 0.88-0.98], number of vitrified oocytes (OR: 1.08, 95% CI: 1.01-1.17) and the day of transfer (OR: 1.97, 95% CI: 1.14-3.42) influenced DR. By recursive partitioning analysis, it can be estimated that more than eight oocytes vitrified are required to improve the outcome (22.6 versus 46.4% DR, respectively). When fewer oocytes are available in women aged >38 years, results are dramatically reduced (12.6 versus 27.5% DR, respectively). Conversely, when >8 oocytes are available, blastocyst culture represents the most efficient policy (62.1% DR; data from one center only). CONCLUSIONS Oocyte vitrification is an efficient and reliable approach, with consistent results between centers and predictable DRs. It should be applied routinely for various indications. A predictive model is proposed to help patient counselling and selection.

Association between human oocyte developmental competence and expression levels of some cumulus genes

At present, oocyte selection is mainly based upon morphological criteria but it is generally acknowledged that its reliability requires further improvement. The aim of this study was to determine whether transcript levels in cumulus cells can provide a useful marker of oocyte developmental competence in vitro. A retrospective study was performed on cumulus cells isolated from 90 oocytes retrieved from 45 patients. Upon fertilization, 35 oocytes originated good-quality embryos and 36 developed into poor-quality embryos, whereas 19 failed to be fertilized. Semi-quantitative measurement of hyaluronic acid synthase 2 (HAS2), gremlin1 (GREM1), and pentraxin 3 (PTX3) mRNAs was performed and data for all genes were obtained from all the samples. Cumulus cells isolated from oocytes that originated high-quality embryos on day 3 of culture had HAS2 and GREM1 transcript levels higher than those detected in cells from oocytes that did not fertilize or developed into poor-quality embryos. No differences were observed in PTX3 levels. Results indicate that the measurement of HAS2 and GREM1 levels in cumulus cells would reliably complement the morphological evaluation providing a useful tool for selecting oocytes with greater chances to be fertilized and develop in vitro.

Sperm banking and rate of assisted reproduction treatment: insights from a 15-year cryopreservation program for male cancer patients.

Semen cryostorage remains the only proven method to preserve fertility in men with cancer. However, the necessity and the justification of banking spermatozoa have been questioned because it has been reported that only a small percentage of patients are referred for assisted reproductive technologies using frozen semen.

Vitamin D Status in Women With Uterine Leiomyomas

CONTEXT Recent in vitro and in vivo experimental evidence supports a role of vitamin D insufficiency as an important factor in the development of uterine leiomyomas. However, epidemiological data supporting this possibility are scanty. OBJECTIVE Our objective was to investigate vitamin D status in women with and without uterine leiomyomas. DESIGN This was a case-control study of women referring to 2 infertility units in Italy. Women were eligible as cases if they were diagnosed with at least 1 uterine leiomyoma with a mean diameter ≥10 mm at transvaginal ultrasound. Each of them was matched to the 2 subsequent women of the same age (±1 year) whose uterus resulted unremarkable at ultrasound. Selected women provided a blood sample for the quantitative detection of 25-hydroxyvitamin D₃ levels. MAIN OUTCOME MEASURE We measured serum concentration of 25-hydroxyvitamin D₃. RESULTS A total of 128 women with leiomyomas and 256 controls were selected. The mean ± SD serum concentration of 25-hydroxyvitamin D3 was significantly lower in affected women compared with controls (18.0 ± 7.7 vs 20.8 ± 11.1 ng/mL respectively, P = .010). The number (proportion) of women with 25-hydroxyvitamin D3 deficiency (ie, <10 ng/mL) in cases and controls was 19 (15%) and 19 (7%), respectively (P = .022). The adjusted odds ratio for the presence of leiomyomas in women with serum levels of 25-hydroxyvitamin D₃ deficiency was 2.4 (95% confidence interval = 1.2-4.9) (P = .016). CONCLUSIONS Vitamin D is an emerging regulator of uterine leiomyoma development. Cohort and interventional studies are pressingly needed to confirm a causal relationship and to investigate the potential therapeutic benefits of vitamin D supplementation.

Vitamin D Deficiency and Infertility: Insights From in vitro Fertilization Cycles

CONTEXT Vitamin D deficiency has been proven to affect fertility in mammals, but data in human is less convincing. In particular, data on in vitro fertilization (IVF), an attractive model to draw information on this topic, are sparse and conflicting. OBJECTIVE Our objective was to investigate IVF outcome in women with deficient 25-hydroxy-vitamin D [25(OH)D] serum levels (<20 ng/mL). DESIGN AND SETTING This prospective cross-sectional study was conducted at the infertility unit of an academic setting. PATIENTS The main inclusion criteria were as follows: (1) indication to IVF, (2) age 18-42 years, (3) BMI 18-25 kg/m(2), (4) adequate ovarian reserve according to Bologna criteria. Eligible women provided a serum sample for 25(OH)D measurement at the time of cycle preparation. Subjects were subsequently excluded if the cycle was cancelled or if the attempt was excessively delayed. INTERVENTION Quantitative detection of serum 25(OH)D. MAIN OUTCOME MEASURE Clinical pregnancy rate. RESULTS The number of recruited women with serum 25(OH)D <20 ng/mL and ≥ 20 ng/mL was 154 and 181, respectively. The clinical pregnancy rates were 20% (30/154) and 31% (56/181), respectively (P = .02); the adjusted odds ratio for clinical pregnancy in women with vitamin D ≥ 20 ng/mL was 2.15 (95% CI: 1.23-3.77). Subgroup analyses showed that the group of women with the highest serum levels (>30 ng/mL) had the highest chances of pregnancy. CONCLUSIONS Vitamin D is an emerging factor influencing female fertility and IVF outcome. Additional studies are pressingly needed to confirm a causal relationship and to investigate the potential therapeutic benefits of vitamin D supplementation.

Cell Lines Derived from Human Parthenogenetic Embryos Can Display Aberrant Centriole Distribution and Altered Expression Levels of Mitotic Spindle Check-point Transcripts

Human parthenogenetic embryos have recently been proposed as an alternative, less controversial source of embryonic stem cell (ESC) lines; however many aspects related to the biology of parthenogenetic embryos and parthenogenetic derived cell lines still need to be elucidated. We present here results on human cell lines (HP1 and HP3) derived from blastocysts obtained by oocyte parthenogenetic activation. Cell lines showed typical ESC morphology, expressed Oct-4, Nanog, Sox-2, Rex-1, alkaline phosphatase, SSEA-4, TRA 1-81 and had high telomerase activity. Expression of genes specific for different embryonic germ layers was detected from HP cells differentiated upon embryoid body (EBs) formation. Furthermore, when cultured in appropriate conditions, HP cell lines were able to differentiate into mature cell types of the neural and hematopoietic lineages. However, the injection of undifferentiated HP cells in immunodeficient mice resulted either in poor differentiation or in tumour formation with the morphological characteristics of myofibrosarcomas. Further analysis of HP cells indicated aberrant levels of molecules related to spindle formation as well as the presence of an abnormal number of centrioles and autophagic activity. Our results confirm and extend the notion that human parthenogenetic stem cells can be derived and can differentiate in mature cell types, but also highlight the possibility that, alteration of the proliferation mechanisms may occur in these cells, suggesting great caution if a therapeutic use of this kind of stem cells is considered.

Risks of conservative management in women with ovarian endometriomas undergoing IVF

BACKGROUND Classical surgical management of endometriotic ovarian cysts using the laparoscopic stripping technique has been recently questioned because of the surgical-related injury to the ovarian reserve. Accordingly, available guidelines suggest that endometriomas with a mean diameter below 4 cm should not be systematically removed before IVF procedures. However, conservative management may have some potential drawbacks and risks. The presence of the endometrioma may theoretically interfere with ovarian responsiveness to hyperstimulation and oocyte competence, the retrieval of the oocytes may be more difficult and risky, the disease may progress during the procedure, pregnancy outcome may be affected and there is the risk of missing occult malignancies with cancer development later in life. In the present review, we aimed at assessing whether these risks do exist and, if so, at estimating their clinical relevance. METHODS We searched PubMed for articles published in the English language between January 1990 and August 2014 that reported on endometriomas and assisted reproductive techniques. Special care was given to studies reporting data purporting to distinguish the effects of ovarian endometriomas per sé from those consequent to surgery for endometriosis or from endometriosis in general. RESULTS Based on the evidence reviewed in the present study, it can be concluded that conservative management may actually expose women to four of the following theoretical risks, i.e. infection of the endometriomas, follicular fluid contamination with the endometrioma content, higher risk of pregnancy complications and cancer development later in life. The first three conditions do not justify surgery because these events are uncommon and the number of women needed to be treated would be exceedingly high and would not justify the costs and risks of the intervention. Albeit also very rare, the possibility of developing ovarian cancer later in life is more troublesome because it is a life-threatening condition. However, this alarmism is supported by only one cohort study and this risk can be effectively prevented by postponing surgery until after the IVF programme is concluded or when women have definitely satisfied their reproductive wishes. CONCLUSION The available evidence on the risks of conservative management does not support systematic surgery before IVF in women with small ovarian endometriomas.

Ovarian endometriomas and oocyte quality: insights from in vitro fertilization cycles

OBJECTIVE To evaluate whether the presence of endometriomas affects ovarian function. DESIGN Prospective cohort study. SETTING Infertility unit of an academic setting. PATIENT(S) Twenty-nine women undergoing an IVF cycle. INTERVENTION(S) Prospective evaluation of women with unoperated unilateral endometriomas undergoing IVF. The affected and contralateral intact gonads were compared in terms of responsiveness and oocyte quality. MAIN OUTCOME MEASURE(S) Oocyte developmental competence. RESULTS Ovarian responsiveness and oocyte quality did not significantly differ between the affected and intact gonads. The number of codominant follicles, the number of oocytes retrieved, and the number of suitable oocytes in affected and intact gonads were 3.7 ± 2.4 and 4.1 ± 1.7, 4.2 ± 3.1 and 4.7 ± 2.5, and 3.1 ± 2.6 and 3.5 ± 2.3, respectively. The number of viable embryos and the number of high-quality embryos were 1.8 ± 2.1 and 1.8 ± 1.4 and 1.0 ± 1.7 and 0.8 ± 0.7, respectively. The fertilization rate in the affected and intact gonads was 64% and 64%, respectively. The cleavage rate was 58% and 51%, respectively. The rate of high-quality embryos was 31% and 21%, respectively. CONCLUSION(S) In women undergoing IVF, the presence of ovarian endometriomas does not affect oocyte developmental competence.

Asymptomatic adenomyosis and embryo implantation in IVF cycles

Research on the effect of adenomyosis on the rate of success of IVF is controversial. Differences in study design, study power, criteria and instrument used to diagnose adenomyosis and choice of controls may explain these discrepancies. To establish whether embryo implantation is impaired in women with adenomyosis, women scheduled for IVF were prospectively evaluated for the presence of adenomyosis and whether this condition affected embryo implantation. Forty-nine women with adenomyosis diagnosed at transvaginal ultrasound with no abnormal uterine bleeding were recruited. They were matched for study period, age, day of embryo transfer and number of transferred embryos to 49 controls without the disease. In women with adenomyosis, 24 out of 76 embryos transferred implanted (32%); this occurred in 16 out of 76 (21%) in unaffected controls. The crude odds ratio of implantation in affected women was 1.73 (95% CI 0.83 to 3.60). The odds ratio adjusted for body mass index (the unique variable found to differ at univariate analysis) was 1.78 (95% CI 0.85 to 3.77). In conclusion, implantation rate is not impaired in asymptomatic women who are diagnosed with adenomyosis at transvaginal sonography. Affected women can be reassured about the effect of this condition on their chances of success.

Confined placental mosaicism at chorionic villous sampling: risk factors and pregnancy outcome

This study aims to investigate the clinical relevance of confined placental mosaicism (CPM) detected at chorionic villous sampling (CVS) and to identify risk factors for this condition.