In Vitro Maturation Cycles Research Articles

Chromosome constitution of human embryos generated after in vitro maturation including 3-isobutyl-1-methylxanthine in the oocyte collection medium.

Do cleavage-stage embryos obtained from oocytes matured in vitro after pre-incubation with a phosphodiesterase inhibitor (IBMX) carry more chromosomal abnormalities than those generated from oocytes matured in vivo? The rate and type of chromosomal abnormalities in normally developing cleavage-stage embryos generated with an in vitro maturation (IVM) system including pre-incubation with IBMX are not different from those observed in supernumerary embryos obtained from oocytes matured in vivo. Very limited information is available about the chromosomal constitution of IVM embryos. Previous studies were carried out using FISH on single biopsied blastomeres or arrested whole embryos and only provided fragmentary information on chromosomal abnormalities in IVM embryos. There is no systematic study of chromosomal abnormalities in all blastomeres of human Day 3 embryos with good morphology. Between July 2012 and December 2012, 16 young (age <35 years old) egg donors underwent 18 IVM cycles for the generation of research embryos. Eighteen embryos developed to Day 3 and were analysed using array comparative genomic hybridization (aCGH). Immature oocytes were retrieved from 2 to 10 mm follicles after mild ovarian stimulation with gonadotrophins but without hCG ovulation trigger. At collection, oocytes were pre-incubated with 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase inhibitor and matured in vitro. After IVM culture, mature oocytes were microinjected with sperm from a single donor. Embryos were cultured to Day 3 after ICSI and all blastomeres of 18 good-morphology embryos were collected individually for aCGH. Oocyte maturation rate in vitro was 50.2% (120/239). The mean fertilization rate was 68.3% (82/120) and 30.5% (25/82) of fertilized oocytes developed into a morphologically good quality embryo on Day 3 after ICSI. Of these, 18 embryos that developed well up to Day 3 were analysed using aCGH. Eighty of the 123 blastomeres analysed showed at least one chromosomal abnormality. Three out of eighteen embryos had completely normal cells. A single embryo carried a meiotic abnormality, 11 embryos were mosaic and three were chaotic. Although the aneuploidy data of this study are too limited to allow statistical analysis, these data are comparable to our own published data on the chromosome constitution of whole day 3 and day 4 embryos after conventional ART. Array CGH technology determines relative quantification of chromosomal domains but does not allow for the visualization of chromosomal rearrangements, assessment of ploidy or detection of uniparental isodisomy. Conclusions drawn on segmental abnormalities should be treated with caution. Although the limited number of embryos analysed here precludes firm conclusions, they provide valuable data on possible causes of the reduced potential of IVM embryos. This is the first study to describe the complete chromosome complement of all single blastomeres of good-morphology day 3 embryos obtained with IVM (including the presence of IBMX in a pre-incubation medium). The results demonstrate that a high proportion of good-morphology embryos are aneuploid and that there is no obvious increase in aneuploidies as a result of IVM which seems to suggest that the reduced efficiency of IVM technology compared with standard IVF may be accounted for by factors other than aneuploidy, such as cytoplasmic defects or reduced endometrial receptivity. This study was funded by the TBM (Applied Biomedical Research with Societal Finality) programme of the IWT (Agency for Innovation through Science and Technology - Flanders, 110680) and by a Methusalem grant of the Vrije Universiteit Brussel. C.S. is a post-doctoral fellow of the Fund for Scientific Research Flanders (FWO - Vlaanderen). K.J. is a PhD student funded by the FWO. The University of Adelaide owns a patent family associated with IVM technologies that is licensed to Cook Medical. R.B.G. and J.G.T. are inventors. The remaining authors have no conflict of interest to declare.

In-vitro maturation of oocytes vs in-vitro fertilization with a gonadotropin-releasing hormone antagonist for women with polycystic ovarian syndrome: can superiority be defined?

ObjectivePatients with polycystic ovarian syndrome (PCOS) are at increased risk of ovarian hyperstimulation syndrome (OHSS) in controlled ovarian hyperstimulation cycles. Interventions to reduce the risk of OHSS in these patients include in-vitro fertilization (IVF) with a gonadotropin-releasing hormone (GnRH) antagonist, and retrieval of immature oocytes followed by in-vitro maturation (IVM). The aim of this study was to compare the outcomes of IVM and IVF-GnRH antagonist protocols in women with PCOS undergoing assisted reproductive technology. Study designRetrospective cohort study. Records of women with PCOS who underwent IVM or IVF-GnRH antagonist protocols between 2010 and 2011 were reviewed. In total, there were 61 IVM cycles and 53 IVF-GnRH antagonist cycles. The treatment protocols were compared in terms of the number of oocytes retrieved, dose of gonadotropin administrated, fertilization rates, quality of embryos, pregnancy, and delivery and abortion rates. ResultsThe number (mean±standard deviation) of mature oocytes did not differ significantly between the two groups (7.11±5.7 vs 8.16±5.07 for the GnRH antagonist group and the IVM group, respectively; p=0.38). The average dose of gonadotropin (1938 IU±838 IU/cycle vs 118±199 IU/cycle; p<0.001), fertilization rate (77% vs 60%; p<0.001) and high-quality embryo rate (58.8% vs 48.3; p<0.001) were significantly higher in the GnRH antagonist group compared with the IVM group. Pregnancy rates (40% vs 25%; p=0.08), livebirth rates per pregnancy (71% vs 53%; p=0.265) and abortion rates (10% vs 27%; p=0.17) were comparable. ConclusionsThe IVM protocol can be an alternative for infertile women with PCOS who wish to prevent the potential adverse effects of gonadotropin treatment. Prospective studies are needed to compare the outcomes of these two treatment protocols.

In vitro maturation rate of immature oocytes according to presence of in vivo matured oocytes

OBJECTIVE: The purpose of this study was to investigate the in vitro maturation rate of immature oocytes according to presence of in vivo mature oocytes in unstimulated in vitro maturation (IVM) cycles. DESIGN: Retrospective study. MATERIALS AND METHODS: 282 patient who underwent unstimulated IVM cycles were included. Without ovarian stimulation using exogenous gonadotrophins, transvaginal ultrasound was performed every 2-3 days from menstrual day 3 to check endometrial thickness and ovarian follicles. Ovulationwas inducedwith urinary or recombinant hCGwhen endometrium reached more than 6mm in thickness, and transvaginal oocyte retrieval was performed 36 hours later. Cumulus cells were removed after oocyte retrieval for assessment of maturity. Among the retrieved oocytes, mature ones were fertilized within 5 hours. The immature ones were subsquently matured in vitro in culture medium for 24-48 hours. After confirmation of maturation, the oocytes were fertilized using intracytoplasmic sperm injection technique. RESULTS: Mean age of the study subjects was 32.4 3.0 years and mean body mass index was 22.7 13.1 Kg/m. Mean blood level of AMH was 10.0 5.2 ng/mL and mean number of total retrieved oocytes was 17.1 8.4. A significant negative correlation was noted between % mature oocytes on retrieval and in vitro maturation rate of immature oocytes (p1⁄40.001, R1⁄4-0.191). In cycles in which mature oocytes were retrieved (n1⁄4125) and not (n1⁄4157), the in vitro maturation rate of immature oocytes were 60.8 19.0% and 66.6 14.7% (P1⁄40.004) respectively. But there were no significant differences in total %mature oocytes (67.0 15.1% vs 66.0 14.3%, p1⁄40.60), fertilization rate (53.3 17.0% vs 52.5 14.1%, p1⁄40.66) and pregnancy rate (37.6% vs 35.0%, p1⁄40.66) between two groups. CONCLUSION: In unstimulated IVM cycles, in vitro maturation rate of immature oocytes was lower when naturally mature oocytes were retrieved. But the presence of in vivo mature oocytes doesn’t affect total oocyte maturation rate, fertilization rate and pregnancy rate.

Fertilization and embryo development with spermatozoa obtained from testicular sperm extraction into oocytes generated from human chorionic gonadotropin-primed in vitro maturation cycles

To evaluate the fertilization rate and embryo development resulting from intracytoplasmic sperm injection (ICSI) of spermatozoa retrieved by testicular sperm extraction (TESE) in hCG-primed in vitro maturation (IVM) cycles. Case-control study. University teaching hospital. Twenty-four IVM cycles were performed in 21 patients (mean age, 32.3 ± 2.4 years) with polycystic ovaries (PCO) whose partners were nonobstructive azoospermic. Twelve cycles where IVM oocytes were also retrieved were compared with a control group consisting of age-matched IVM cycles with ICSI using ejaculated spermatozoa (n = 12). In vitro maturation treatment with TESE sperm. Fertilization and embryo development between sibling oocytes matured in vivo and in vitro. Eight singleton pregnancies and one twin pregnancy were obtained after ET (9/24, 37.5%). In the 12 IVM cycles where in vivo-matured oocytes were also obtained, the fertilization rate after TESE-ICSI was significantly higher in in vivo-matured oocytes than in sibling in vitro-matured oocytes (84.2% vs. 53.2%). The proportion of good quality embryos was also higher (63.5% vs. 40.2%). In the control group of cycles with ejaculated spermatozoa, there was no difference in fertilization rates between sibling oocytes matured in vivo and in vitro (84.6% vs. 79.6%). Our results suggest that IVM of immature oocytes combined with TESE-ICSI is an option for couples with PCO and azoospermia. However, there are lower fertilization and good quality embryo rates achieved when TESE-ICSI was done with in vitro-matured oocytes. Additional studies are necessary to determine the role of this treatment combination.

What is the optimal threshold of serum Anti-Müllerian hormone (AMH) necessary for IVM treatments?

To assesse circulating levels of Anti-Müllerian hormone (AMH) as a predictor of oocyte number and their potential to mature in vitro in both normo-ovulatory (NO) women and in women with Polycystic Ovary Syndrome (PCOS) undergoing in vitro maturation (IVM) treatments. We prospectively studied NO women and women diagnosed with PCOS, (age range 21-39 years) underwent IVM treatments at our center. Serum AMH levels were quantified before each cycle and correlated to oocytes number, maturation and fertilization during in vitro maturation. 104 NO and 30 PCOS IVM cycles were followed with retrieval of a total of 672 and 491 oocytes, respectively. In NO women, the serum AMH level positively correlated with the number of oocytes retrieved, (R = 0.6; P <0.0001) the number of M2 oocytes at 24 and 48 h (R = 0.4; P <0.01; R = 0.26 p < 0.007, respectively) and with the total number of M2 oocytes (R = 0.47; P < 0.0001). In the PCOS group, the serum AMH level positively correlated only with the number of oocytes retrieved (R = 0.43; P <0.03). Receiver operating characteristic (ROC) analyses showed that a cutoff AMH level of 1.56 (ng/ml) could identify patients with 5 or more oocytes at OPU with a sensitivity of 83 % and a specificity of 75 %. An AMH level of 1.63 (ng/ml) was the threshold for 5 or more matured oocytes (sensitivity = 81 %, specificity = 53 %). Serum AMH may be used as a marker to identify candidates for IVM treatment in both NO and PCOS women.

A prediction model to select PCOS patients suitable for IVM treatment based on anti-Mullerian hormone and antral follicle count

Which baseline patient characteristics can help assisted reproductive technology practitioners to identify patients who are suitable for in-vitro maturation (IVM) treatment? In patients with polycystic ovary syndrome (PCOS) who undergo oocyte IVM in a non-hCG-triggered system, circulating anti-Müllerian hormone (AMH), antral follicle count (AFC) and total testosterone are independently related to the number of immature oocytes and hold promise as outcome predictors to guide the patient selection process for IVM. Patient selection criteria for IVM treatment have been described in normo-ovulatory patients, although patients with PCOS constitute the major target population for IVM. With this study, we assessed the independent predictive value of clinical and endocrine parameters that are related to oocyte yield in patients with PCOS undergoing IVM. Cohort study involving 124 consecutive patients with PCOS undergoing IVM whose data were prospectively collected. Enrolment took place between January 2010 and January 2012. Only data relating to the first IVM cycle of each patient were included. Patients with PCOS underwent oocyte retrieval for IVM after minimal gonadotrophin stimulation and no hCG trigger. Correlation coefficients were calculated to investigate which parameters are related to immature oocyte yield (patient's age, BMI, baseline hormonal profile and AMH, AFC). The independence of predictive parameters was tested using multivariate linear regression analysis. Finally, multivariate receiver operating characteristic (ROC) analyses for cumulus oocyte complexes (COC) yield were performed to assess the efficiency of the prediction model to select suitable candidates for IVM. Using multivariate regression analysis, circulating baseline AMH, AFC and baseline total testosterone serum concentration were incorporated into a model to predict the number of COC retrieved in an IVM cycle, with unstandardized coefficients [95% confidence interval (CI)] of 0.03 (0.02-0.03) (P < 0.001), 0.012 (0.008-0.017) (P < 0.001) and 0.37 (0.18-0.57) (P < 0.001), respectively. Logistic regression analysis shows that a prediction model based on AMH and AFC, with unstandardized coefficients (95% CI) of 0.148 (0.03-0.25) (P < 0.001) and 0.034 (-0.003-0.07) (P = 0.025), respectively, is a useful patient selection tool to predict the probability to yield at least eight COCs for IVM in patients with PCOS. In this population, patients with at least eight COC available for IVM have a statistically higher number of embryos of good morphological quality (2.9 ± 2.3; 0.9 ± 0.9; P < 0.001) and cumulative ongoing pregnancy rate [30.4% (24 out of 79); 11% (5 out of 45); P = 0.01] when compared with patients with less than eight COC. ROC curve analysis showed that this prediction model has an area under the curve of 0.7864 (95% CI = 0.6997-0.8732) for the prediction of oocyte yield in IVM. The proposed model has been constructed based on a genuine IVM system, i.e. no hCG trigger was given and none of the oocytes matured in vivo. However, other variables, such as needle type, aspiration technique and whether or not hCG-triggering is used, should be considered as confounding factors. The results of this study have to be confirmed using a second independent validation sample. The proposed model could be applied to patients with PCOS after confirmation through a further validation study. This study was supported by a research grant by the Institute for the Promotion of Innovation by Science and Technology in Flanders, Project number IWT 070719.

Comparison of the obstetric and perinatal outcomes of children conceived from in vitro or in vivo matured oocytes in in vitro maturation treatments with births from conventional ICSI cycles

Are the obstetric and perinatal outcomes of deliveries following in vitro maturation (IVM) cycles different from births generated from controlled ovarian stimulation (COS) cycles? The obstetric and perinatal outcomes of births from IVM cycles are comparable with those of ICSI treatments, including the incidence of major and minor abnormalities. Only few and numerically small reports on the health of IVM children are currently available. Retrospective cohort study involving 196 babies born from IVM cycles carried out with different priming regimens. Of these children, 79 developed from oocytes matured in vitro after 30 h of culture, while 104 originated from oocytes found mature and inseminated on the day of recovery. Thirteen babies were obtained from embryos developed from both types of oocytes. Data of these births were compared with those of 194 children born from COS ICSI cycles performed during the same period (March 2004 to December 2011). IVM cycles were done in the absence of gonadotrophin administration or with FSH and/or HCG priming. All oocytes were inseminated by microinjection. ICSI and ICSI cycles were chosen as a control group to exclude possible influences of the insemination technique. Couples in which maternal age was >39 years or affected by azoospermia were excluded to rule out major parental effects. In single births, gestational age at delivery was comparable, but birthweight was significantly higher (P = 0.009) in children from IVM cycles (3091 ± 669 versus 3269 ± 619 g). In a separate analysis of the IVM group, comparing singleton births derived with certainty from oocytes matured in vitro (n = 71) or in vivo (n = 74), no statistically significant differences were observed in terms of birthweight (3311 ± 637 versus 3194 ± 574 g, respectively) and gestational age (38.9 ± 2.4 versus 38.4 ± 2.1 weeks, respectively). In twin births, gestational age was lower in IVM cycles, while weight at birth was comparable (ICSI, 2432 ± 540 g; IVM, 2311 ± 577 g). In single births, major and minor abnormalities were 2 (1.4%) and 6 (4.1%) in the ICSI group and 0 (0.0%) and 8 (5.2%) in the IVM category, respectively. In twin children, major and minor abnormalities were 1 (2.2%) and 2 (4.3%) in ICSI babies and 0 (0.0%) and 2 (4.6%) in IVM cycles, respectively. The study is the largest conducted so far. Nevertheless, it is limited by its retrospective nature and the fact that most births of IVM treatments derived from oocytes found mature at recovery in cycles primed with HCG. A more comprehensive appraisal of the health status of IVM children will demand larger prospective studies. The study is consistent with previous reports suggesting a possible role of standard ovarian stimulation in determining a reduced birthweight in children born from COS cycles. No external funding was sought to support this work. The authors have no conflicts of interest to declare. Not applicable.

PCOS patients can benefit from in vitro maturation (IVM) of oocytes

Our aim was to compare treatment outcome following in vitro maturation (IVM) compared with IVF in patients with polycystic ovarian syndrome (PCOS). Retrospective evaluation of treatment in women with PCOS who underwent IVM (108) and IVF (108). We found a significant difference in outcome between IVM and IVF, with an increase in the number of mature oocytes derived (10.5 ± 6.5 vs. 15.3 ± 8.8, p<0.0001) and the cleavage rate (92.4 ± 13.0 vs. 95.2 ± 11.7, p=0.03) in IVM cycles. Due to the lower implantation rate (16.1% vs. 21.6%, p=0.07) we tend to transfer more embryos in the IVM group (3.4 ± 0.8 vs. 2.8 ± 1.0, p<0.0001), but the multiple pregnancy rate in that group was not higher. Importantly, the delivery rate was similar in both groups (26.8% vs. 25%). We also report a yearly change in the success rate of IVM during this period. IVM treatment for PCOS patients may be a valid alternative treatment to IVF with the advantage of eliminating the risk of OHSS and reducing the cost of medication, whilst maintaining high clinical pregnancy rate.

Developmental capacity of in vitro–matured human oocytes retrieved from polycystic ovary syndrome ovaries containing no follicles larger than 6 mm

To test the developmental competence of oocytes in a nonhCG-triggered in vitro maturation (IVM) system when oocyte-cumulus complexes (OCC) are retrieved from antral follicles with a diameter of <6 mm. Prospective cohort study. Tertiary university-based referral center. From January 2010 to September 2011, 121 patients with polycystic ovaries/polycystic ovary syndrome underwent 239 IVM cycles in total. In 58 of these cycles (44 patients), all antral follicles had a diameter of <6 mm on the day of oocyte retrieval. NonhCG-triggered IVM of oocytes, fresh or vitrified/warmed embryo transfer (ET). Oocyte diameter, maturation rate, fertilization rate, embryo development and morphology, implantation rate, clinical pregnancy rate, ongoing pregnancy rate. Oocyte retrieval yielded 16.7 OCC/cycle, and 50.8% of oocytes completed IVM. The mean oocyte diameter increased from 108.8 ± 4.3 μm to 111.9 ± 4.1 μm after IVM. Mean fertilization rate was 63.7%, and 45.4% of 2-pronuclei oocytes developed into a morphologically good-quality embryo on day 3 after intracytoplasmic sperm injection. Fresh ET resulted in two ongoing pregnancies (2/37; 5.4%). Deferred vitrified-warmed ET led to an ongoing pregnancy rate of 34.6% (9/24). Three healthy babies were born and eight pregnancies were still ongoing. Oocytes retrieved from follicles with a diameter of <6 mm grow during a 40-hour IVM culture can acquire full competence in vitro, as illustrated by their development into healthy offspring. Endometrial quality appears to be a crucial determinant of pregnancy after nonhCG-triggered IVM.

Successful pregnancy following a novel endometrial preparation in a PCOS patient undergoing IVM: a case report

In-vitro fertilization (IVF) relies heavily on controlled ovarian stimulation to increase the number of available oocytes and embryos, and therefore the pregnancy rate. However, the side effects, such as nausea, mood swings, menopausal symptoms, ovarian hyperstimulation syndrome (OHSS), and potential cancer risk of ovarian stimulation may not be avoided. Therefore, the recovery of immature oocytes followed by in-vitro maturation (IVM) and fertilization is an attractive alternative to avoid the side effects of using gonadotrophin stimulation. In IVM, immature oocytes are retrieved from unstimulated or minimally stimulated cycles, then cultured using IVM culture media for maturation in vitro [1]. In comparison with conventional IVF, IVM technology is generally associated with the absence of OHSS, lower cost, and fewer complications. However, the efficiency of existing IVM technique is suboptimal because embryos generated from IVM exhibit frequent cleavage blocks and low implantation rate [2]. Women with polycystic ovarian syndrome (PCOS) are considered as the target population for this approach. This group of patients is extremely sensitive to ovarian stimulation and have a high potential for developing of OHSS [3]. In recent years, many improvements have been made in clinical and laboratory aspects of IVM treatment which have resulted in better pregnancy outcomes [4]. It was shown that pretreatment with FSH in the early days of folliculogenesis phase may increase the number of retrieved immature oocytes and/or maturation potential [5, 6]. In addition, hCG priming promotes some GV oocytes to reach the MI stage and increases the maturation rate of immature oocytes in vitro setting. Despite increased pregnancy rates following IVM in recent years, implantation rates are still lower than conventional IVF cycles. One possible explanation may be related to insufficient endometrial preparation during IVM cycles before ET [4, 7]. Here, we aim to report a novel endometrial preparation method in a PCOS case undergoing IVM. Case report A 33 years old patient with history of primary infertility for 3 years was referred to our program for infertility treatment. She was diagnosed as PCOS according to Rotterdam criteria [8] with history of acne, hirsutism and oligomenorrhea. The clinical profile showed that she had BMI = 28, LH = 4, FSH = 1.5, LH/FSH > 2. The patient had history of both clomiphen failure and metformin administration for more than 6 months. She also underwent laparoscopy and ovarian cauterization approximately 1 year before. Infertility work up for the husband showed abnormal sperm analysis including a count of 28 millions/ml, <20% progressive motility and <14% normal morphology according to WHO criteria. A recent varicoceletomy did not improve semen quality for her husband. In her first treatment cycle and due to a high response to the antagonist protocol and to avoid the risk of severe OHSS, it was decided to cancel the cycle, and to continue the treatment using IVM techniques. After progesterone withdrawal bleeding, FSH priming was delayed until 13th day of the cycle when endometrial thickness reached 7 mm. At that time, FSH priming started, 75 IU/day of FSH (Serono, Aubnne, Switzerland) was injected for 3 days, followed by an injection of 10,000 IU hCG (Pregnyl, Organon, Netherlands). Follicular diameters were less than 10 mm at the time of hCG injection. On the day of hCG injection, both endometrial thickness and level of estradiol were measured. Oocytes were retrieved 36 h after hCG injection using a 19-gauge aspiration needle under transvaginal ultrasound guidance. A total of 10 immature oocytes were retrieved, including 3 at GV stage, 5 at MI stage, and 2 were degenerated. Immature oocytes were transferred to IVM culture media (SAGE Co., CT) for 28 h, when 5 oocytes reached MII stage. The rest of oocytes were arrested. The mature oocytes were microinjected with husbands’ sperm and normal fertilization was confirmed when two polar bodies with two pronuclei were observed 18–20 h after insemination. A total of 3 oocytes were fertilized and 3 four cells embryos (grade B) were transferred 2 days after microinjection using ET catheter (Labotect, Gottingen, Germany). Lutheal phase support was done with 100 mg progesterone in oil (Aburaihan Co, Tehran, Iran) injected daily. Estradiol valerate (Aburaihan C,Tehran, Iran) was taken orally at the dose of 6 mg/day from day of oocytes retrieval, and continued until confirmation of fetal heart by sonography. βhCG was measured 14 days after ET, which was reported positive (>180 IU/L). A healthy baby girl was born at 39 weeks of pregnancy by cesarean section. The baby weigh at the time of birth was 3,500 g with the apgar of 9/10.

Human oocyte maturation in vitro

Oocytes from medium-sized antral follicles have already completed their growth phase and, if released from the follicular environment and cultured in vitro, are able to resume the meiotic process and mature. However, in vitro maturation (IVM) does not entirely support all the nuclear and cytoplasmic changes that occur physiologically as an effect of the ovulatory stimulus. Regardless, oocyte IVM is widely applied for the breeding of agriculturally important species. In assisted reproduction technology, IVM has been proposed as an alternative treatment to circumvent the drawbacks of standard ovarian stimulation regimens. Initially introduced to eliminate the risks of ovarian hyperstimulation syndrome afflicting women presenting with polycystic ovaries, subsequently IVM has been suggested to represent an additional approach suitable also for normovulatory patients. So far, in children born from IVM cycles, no doubts of an increased incidence of congenital abnormalities have been raised. Many more births would be achieved if novel IVM systems, currently dominated by empiricism, could be conceived according to more physiological criteria. Recent findings shedding new light on the control of meiotic progression, the support of cumulus cells to the oocyte cellular reorganization occurring during maturation, and the modulation of the stimulus that promotes oocyte maturation downstream the mid-cycle gonadotropin signal are likely to provide crucial hints for the development of more efficient IVM systems.

Impaired mitochondrial function in murine oocytes is associated with controlled ovarian hyperstimulationand in vitro maturation

The present study was designed to determine whether controlled ovarian hyperstimulation (COH) and in vitro maturation (IVM), two common clinical procedures in human IVF treatment, have an impact on mitochondrial DNA (mtDNA) copy number and mitochondrial function in oocytes. Matured mouse oocytes recovered following COH, IVM and natural cycles (NC), which simulated those treatments in human clinic IVF treatment. The copies of mtDNA, the activity of mitochondria as determined by inner mitochondrial membrane potential and oocyte adenosine trisphosphate (ATP) content, pattern of mitochondrial distribution, reactive oxygen species (ROS) levels and the integrity of the cytoskeleton were evaluated in oocytes. Significant differences were detected between COH and NC groups in all measures, except the pattern of mitochondrial distribution and ROS levels. There were also significant differences detected between IVM and NC treatment groups in the copies of mitochondrial DNA, the level of ROS and the integrity of the cytoskeleton in oocytes. In conclusion, the results of this investigation indicate that non-physiological COH and IVM treatments inhibit mtDNA replication, alter mitochondrial function and increase the percentage of abnormal cytoskeleton and ROS production. Damage related to the mitochondria may partly explain the low efficiency of IVF and high rate of embryonic loss associated with these clinical procedures.

How old is too old for in vitro maturation (IVM) treatment?

ObjectiveFertility declines with age. A study of the outcomes of in vitro maturation (IVM) in women of different ages has not been reported to date. The aim of our study was to evaluate the impact of age on treatment response and on pregnancy rates after IVM treatment. Study designWe reviewed the data of all IVM cycles and recorded the total number of oocytes, total metaphase II (MII) oocytes, the number and quality of embryos as well as pregnancy rates. The main outcome measures were number and maturity of retrieved oocytes and pregnancy rates. ResultsCompared to cycles in women ≤25 years, the number of oocytes and total MII oocytes were significantly lower in women over 40 years. The number of good quality embryos in women ≤25 years (3.6±1.7) was higher than in women aged 40 years (0.8±1.2, P<0.0001). The ongoing pregnancy rates in women aged 20–25 years were 36.8%, 26–35 years were 30.0% and in those 36–39 years were 31.9%. No clinical pregnancy was observed in women over the age of 40. ConclusionWomen over 40 years are poor candidates for IVM treatment.

Novel protocol for scheduling oocyte retrieval in IVM cycles in PCOS patients: a case series

In-vitro maturation (IVM) is associated with a longer egg-collection procedure in the operating room and a longer oocyte-handling time in the IVF laboratory than standard IVF. Hence, if the designated day of oocyte retrieval could be planned in advance, the workload pressure on that specific day can be planned in advance. This study presents a simple method for advance scheduling of IVM in patients with polycystic ovary syndrome (PCOS). A fixed protocol of oral contraceptive pill administered prior to gonadotrophin priming and based on the days of the week enable the exact dating of the oocyte retrieval day, thereby increasing patient convenience and improving control of the IVF-unit workload. This protocol was compared with immediate-start IVM and resulted in a similar pregnancy rate (43.8% and 40.0% per cycle, respectively).

Clinical outcome of non–hCG-primed oocyte in vitro maturation treatment in patients with polycystic ovaries and polycystic ovary syndrome

To compare clinical outcomes of fresh embryo transfer (ET) and vitrified-warmed ET in an artificial endometrial priming cycle in patients with polycystic ovaries (PCO) or polycystic ovary syndrome (PCOS) who underwent oocyte in vitro maturation (IVM) in non-hCG-primed cycles. Prospective cohort study. University-based tertiary referral center. Thirty-nine consecutive patients <37 years old with PCO or PCOS, who underwent 73 cycles of immature oocyte retrieval. Immature oocyte collection after ovarian stimulation with a cumulative dose of 450 IU uFSH or highly purified hMG, but without hCG priming. IVM of oocytes followed by ET if endometrium thickness ≥ 6 mm. Embryo vitrification at the cleavage stage. ET in an artificial cycle. Implantation rate (IR) and clinical pregnancy rate (CPR). Fresh ET after IVM resulted in an IR of 6.9% (5/72) per ET and a CPR of 9.4% (5/53). ET of vitrified-warmed IVM embryos in an artificial cycle resulted in significantly better outcomes (IR 21.9% [7/32] and CPR 31.8% [7/22] per ET). A non-hCG-primed IVM system in PCO or PCOS performs poorly when embryos are transfered in a fresh cycle. Transfer of vitrified-warmed IVM embryos in an artificial cycle leads to significantly improved clinical outcomes. These data illustrate that IVM embryos in PCO or PCOS have good survival rates and suggest that hCG may be needed to support endometrial receptivity in the fresh IVM cycle.

Priming in vitro maturation cycles with gonadotropins: salvage treatment for nonresponding patients

To compare the treatment outcomes in in vitro maturation (IVM) cycles primed with human menopausal gonadotropin with those for pure IVM cycles in patients with polycystic ovary syndrome. Prospective observational. University-based tertiary medical center. Patients undergoing IVM cycles (primed IVM, 47; pure IVM, 118). IVM treatment with and without human menopausal gonadotropin stimulation. Pregnancy rates. The clinical pregnancy rate demonstrated a tendency toward improvement in the primed IVM group (53.1% vs. 43.6%, 20.1% vs. 14.0% and 40.4% vs. 30.8%, [corrected] respectively) with better implantation and delivery rates (20.1% versus 14.4%; 95% confidence intervals 1.0-3.06 and 40.4% versus 24.6%; 95% confidence intervals 0.1-0.8, respectively). We found no significant difference in pure IVM compared with primed IVM in the number of eggs collected, size of leading follicle, fertility rate, cleavage rate, and the number of embryos transferred. Total mature eggs and maturation rate were significantly higher in the group of pure IVM (11 ± 2.1 versus 8.7 ± 0.5 and 68.5% ± 17.5% versus 60.9% ± 0.4%, respectively). Importantly, the endometrial thickness was significantly improved in primed IVM cycles (7.9 ± 1.9 mm versus 7.1 ± 0.8 mm), possibly leading to better implantation and pregnancy rates. Patients who fail to demonstrate endometrial or follicular growth during IVM cycles may benefit from gonadotropin priming during the same cycle.

Comparison of fertilization and embryonic development in sibling in vivo matured oocytes retrieved from different sizes follicles from in vitro maturation cycles

To examine the fertilization and developmental potential of sibling mature oocytes collected from different follicle sizes on day of retrieval in in vitro maturation (IVM) cycles. Two hundred thirty eight hCG-primed IVM cycles were performed in 213 patients with polycystic ovaries. If sibling mature oocytes were retrieved on day of collection, they were divided into two groups, Group 1 (n = 78): M-II oocytes obtained from follicles size 10-14 mm; Group 2 (n = 192): M-II oocytes obtained from follicles size <10 mm. Of the 238 cycles, 63 cycles had more than one M-II oocytes retrieved (total M-II oocytes = 270) both from Groups 1 and 2. There were no significant differences between the two groups for oocyte diameter (117.2 mm vs. 116.9 mm), fertilization (79.5% vs. 72.4%) and good quality embryo on day 3 (66.1% vs. 56.8%). The M-II oocytes retrieved from the cohort of follicles (<10 mm diameter) can produce the same quality of embryos as that from large follicles, likely contributing to improve the clinical outcome.

Ovarian Hyperstimulation Syndrome Prevention Strategies: In Vitro Maturation

The only reliable way to eliminate the risk of ovarian hyperstimulation syndrome (OHSS) is complete avoidance of gonadotropin ovarian stimulation. It could be argued that in vitro maturation (IVM) of oocytes represents the most effective strategy to prevent OHSS. IVM has been an established treatment option in many centers worldwide for over a decade. The use of IVM and natural cycle in vitro fertilization (IVF) combined with IVM can result in clinical pregnancy rates that compare to those obtained with conventional IVF. The obstetric and perinatal outcomes of IVM pregnancies are similar to those conceived from stimulated IVF or spontaneous conceptions. To date, more than a thousand healthy infants have been born without an increase in fetal abnormalities. Although IVM may not replace standard IVF, it plays an increasingly important role in assisted reproductive technology, especially in the settings of high responders and those patients at risk of OHSS.

Fertilization outcome, embryo development and birth after unstimulated IVM.

A common observation in oocyte in vitro maturation (IVM) cycles is poor embryo quality. However, no study was dedicated to assess zygote and early cleavage embryo quality in IVM cycles. The objective of this study is to analyze fertilization outcome, embryo development and the resulting pregnancy and births in unstimulated IVM cycles. IVM oocytes were collected 36 h post hCG and matured in vitro for 28-30 h. All oocytes were inseminated by ICSI. Resulting zygotes and embryos were assessed on day-1, 2 and 3, when transfers were made. The overall oocyte maturation and fertilization rates were 63% and 62%, respectively. Abnormal fertilization rate was 27%. [corrected] Ninety five and 14.6% of the 2Pn zygotes reached the 2-cell and 8-cell stage at day-2 and day-3, respectively. Embryo quality assessment on day-3 at transfer revealed that only 9% of the embryos were of very good quality. Most embryos showed developmental delay. An average of 3.29 embryos were transferred per patient resulting in implantation and clinical gestation rates of 16% and 32%, respectively. Overall 14 healthy babies were born and there is one ongoing pregnancy. Results show a significant rate of abnormal fertilization and poor embryo quality after IVM, which is reflected in a higher than average number of embryos being transferred. However, pregnancy, implantation and birth rates are reasonably high and allow us to consider IVM a valuable approach for the treatment of infertility in PCO or PCOS patients.

The clinical outcomes of vitrified blastocysts on in vitro maturation cycles

OBJECTIVE: Even if cryopreservation is now a routine procedure in human IVF, there are a few reports that blastocysts generated from in vitro maturation (IVM) were frozen using vilification. The purpose of this study is to report production and cryopreservation of blastocysts from IVM. DESIGN: A retrospective study. MATERIALS AND METHODS: The blastocysts produced from IVM cycles were frozen. The embryos were vitrified using an EM grid following artificial shrinkage. The vitrification solution was composed of DPBS containing 20% (v/v) SSS, 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, and 0.3 M sucrose and vitrification was prepared according to the previously described method (Son et al., 2003). Thawing of the vitrified embryos was carried out using a two-step cryoprotectant dilution method (Lee et al., 2006). Thawed blastocysts were cultured for overnight, and then these were observed and transferred. RESULTS: Total 874 blastocysts (240 cycles, 231 patients) were cryopreserved on IVM cycles from 2004 to 2009. Among these, 274 blastocysts (104 cycles, 94 patients) were thawed and transferred. It showed a high rate of post-thawing survival (94%, 259/274). After the transfer of 254 embryos (2 ± 0.82), 44 women (43%) became clinically pregnant and 55 embryos (22%) implanted. Among these women, 21 pregnancies ended in miscarriage, 18 healthy babies were born in 16 deliveries, and 8 patients' pregnancies are currently ongoing. CONCLUSION: The blastocysts produced from IVM could be safely cryopreserved by vitrification using an EM grid. Thus, vitrification is an effective and practical method for cryopreservation in IVM.