Immature Oocyte Retrieval Research Articles

Cryopreservation of in vitro matured oocytes after ex vivo oocyte retrieval from gynecologic cancer patients undergoing radical surgery.

ObjectiveThe aim of this study was to report a case series of in vitro matured (IVM) oocyte freezing in gynecologic cancer patients undergoing radical surgery under time constraints as an option for fertility preservation (FP).MethodsCase series report. University-based in vitro fertilization center. Six gynecologic cancer patients who were scheduled to undergo radical surgery the next day were referred for FP. The patients had endometrial (n=2), ovarian (n=3), and double primary endometrial and ovarian (n=1) cancer. Ex vivo retrieval of immature oocytes from macroscopically normal ovarian tissue was followed by mature oocyte freezing after IVM or embryo freezing with intracytoplasmic sperm injection.ResultsA total of 53 oocytes were retrieved from five patients, with a mean of 10.6 oocytes per patient. After IVM, a total of 36 mature oocytes were obtained, demonstrating a 67.9% maturation rate. With regard to the ovarian cancer patients, seven IVM oocytes were frozen from patient 3, who had stage IC cancer, whereas one IVM oocyte was frozen from patient 4, who had stage IV cancer despite being of a similar age. With regard to the endometrial cancer patients, 15 IVM oocytes from patient 1 were frozen. Five embryos were frozen after the fertilization of IVM oocytes from patient 6.ConclusionImmature oocytes can be successfully retrieved ex vivo from macroscopically normal ovarian tissue before radical surgery. IVM oocyte freezing provides a possible FP option in patients with advanced-stage endometrial or ovarian cancer without the risk of cancer cell spillage or time delays.

Fertility preservation and breast cancer: a review.

Breast cancer is the most common malignancy in women, and its incidence increases with age, with the majority of patients diagnosed after menopause. However, in 15–25% of cases, patients are premenopausal at the time of diagnosis, and about 7% of them are below the age of 40. Therefore, a considerable amount of young women are diagnosed with breast cancer during their reproductive life. Within this group, most cancer cases require cytotoxic chemotherapy and/or hormone therapy, which are responsible for a decrease in the patients’ reproductive function, along with their age. The efficacy of such treatments, among other factors, has led to a high five-year-survival rate, which results in an increasing number of young women who survive breast cancer before having fulfilled their reproductive wishes, especially considering the current trend to delay pregnancy until the late 30s or early 40s in developed countries. The combination of these factors justifies the importance of fertility preservation and reproductive counselling at the time of breast cancer diagnosis in young women.A wide range of fertility preservation techniques has been developed, such as ovarian suppression, oocyte and embryo cryopreservation, immature oocyte retrieval and in vitro maturation, and ovarian tissue cryopreservation.Early counselling and referral of these patients to fertility specialists are fundamental factors in order to maximise their chances of pregnancy. This review aims to update the knowledge about the influence of breast cancer in fertility, the influence of pregnancy and fertility preservation techniques in breast cancer patients and assessment of ovarian reserve for a better treatment choice. A special section dedicated to BRCA-mutation carriers has been included because of their specific features.A comprehensive literature search has been conducted, including publications from the last five years.

GnRH-agonist Ovulation Induction May Be Insufficient to Trigger Follicular Maturation in the Patient with Hypothalamic Hypogonadism

Patients with hypothalamic amenorrhea are at increased risk for developing ovarian hyperstimulation syndrome (OHSS) during controlled ovarian hyperstimulation (COH) cycles1,2 and have a higher risk of retrieval of immature oocytes whose use for subsequent IVF would only be possible following in vitro maturation (IVM)1,2. Consequently, GnRH antagonist IVF cycles with GnRH-agonist (GnRH-a) ovulation induction are preferred for abrogating OHSS risk3 in these patients. However, use of GnRH-a for ovulation induction can lead to empty follicles or immature oocytes but the reported incidence of this is only 1.4%4 and it has not been looked at in the context of patients with hypothalamic amenorrhea.

Follicular fluid lipid fingerprinting from women with PCOS and hyper response during IVF treatment.

Polycystic ovary syndrome (PCOS) is an endocrine-metabolic disorder that leads to lower natural reproductive potential and presents a challenge for assisted reproductive medicine because patients may exhibit immature oocyte retrieval and a higher risk of ovarian hyper stimulation syndrome during in vitro fertilization (IVF) treatment. This study aimed to identify potential lipid biomarkers for women with PCOS and a hyper response to controlled ovarian stimulation. Follicular fluid samples were collected from patients who underwent IVF, including normal responder women who became pregnant (control group, n = 11), women with PCOS and a hyper response to gonadotropins (PCOS group, n = 7) and women with only hyper response to gonadotropins (HR group, n = 7). A lipidomic analysis was performed by electrospray ionization mass spectrometry, and candidate biomarkers were analyzed by tandem mass spectrometry experiment. The lipid profiles indicated particularities related to differences in phosphatidylcholine (PCOS and HR), phosphatidylserine, phosphatydilinositol and phosphatidylglycerol (control), sphingolipids (PCOS) and phosphatidylethanolamine (control and HR). These findings contribute to the understanding of the molecular mechanisms associated with lipid metabolism in the PCOS-related hyper response, and strongly suggest that these lipids may be useful as biomarkers, leading to the development of more individualized treatment for pregnancy outcome.

IVM oocyte cryopreservation after ex-vivo oocyte retrieval in gynecologic cancer patients undergoing radical surgery

In case of advanced gynecologic malignancy, radical surgery is necessary and reproductive organs such as uterus and ovaries could not be spared. Here we report novel approach for fertility preservation (FP) by means of ex-vivo retrieval of immature oocytes from macroscopically normal ovarian tissue harvested during radical surgery in endometrial or ovarian cancer patients with advanced stage.

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.

Growth Development of French Children Born after In Vitro Maturation

BackgroundSeveral lines of evidence indicate that immature oocyte retrieval and subsequent in vitro maturation (IVM) without ovarian stimulation may be a reliable option in assisted reproductive technologies (ART). However, few outcome data are available for children born following this technique.ObjectiveWe assessed height and weight development of French children conceived after IVM.MethodsAll children conceived after IVM at Antoine Beclere Hospital (Clamart, France) and born between June 2003 and October 2008 (n = 38) were included in a prospective cohort study and compared with a control group of children conceived by ICSI without IVM, matched for maternal age, gestational age and singleton/twin pregnancies. Follow-up included clinical examination at one year and a questionnaire completed by parents when the children were two years old (97% follow-up rate).ResultsNo statistical differences between IVM and control groups were found for boys. Mean weight, height and head circumference at birth were significantly greater for IVM than for ICSI girls (3.236 kg vs 2.701 kg (p = 0.03); 49 cm vs 47 cm (p = 0.01) and 34 cm vs 33 cm (p = 0.04), respectively). At one year, IVM girls remained heavier (mean weight 10.2 kg vs 8.6 kg (p = 0.001)) and taller (76 cm vs 73 cm (p = 0.03)), and there was a two-point difference in BMI between the two groups of girls (18 vs 16 (p = 0.01)).ConclusionOur results in girls born after IVM should be interpreted with caution. It remains unclear whether the observed sexual dimorphism is due to IVM technology or to maternal characteristics such as underlying infertility in patients with polycystic ovary syndrome (PCOS). Further monitoring of the outcomes of these infants is required.

The effect of ovarian puncture on the endocrine profile of PCOS patients who undergo IVM

BackgroundTo examine whether ovarian puncture for immature oocyte retrieval and in-vitro maturation (IVM) has an effect on the endocrine profile of patients with polycystic ovary syndrome (PCOS).MethodsTwenty-two consecutive patients with PCOS undergoing IVM treatment were included. Serum anti-Müllerian hormone (AMH), sex hormone-binding globulin (SHBG), total testosterone (TT) and luteinized hormone (LH) levels were analyzed at the start of the cycle, on the day of immature oocyte retrieval (OR) and at fixed intervals thereafter, for up to three months after OR.ResultsFive days after OR circulating AMH, TT, calculated free testosterone (FTc), and LH levels were significantly reduced and circulating SHBG was significantly increased. Two weeks after OR, TT, FTc and LH remained reduced, whereas circulating AMH and SHBG levels recovered to pre-puncture values. Three months after OR, all circulating hormone levels had recovered to baseline values.ConclusionOvarian puncture for the retrieval of immature oocytes and IVM in patients with PCOS has a significant impact on the ovarian endocrine profile, but this impact is brief and transient.

In vitro maturation of immature human oocytes for clinical application.

Meiotic progression in the oocyte is defined as oocyte maturation from reinitiation of the first meiotic division from the germinal vesicle (GV) stage to the metaphase-II (M-II) stage (Fig. 1), (Cha and Chian, Hum Reprod Update 4:103-120, 1998). Priming with FSH or HCG prior to immature oocyte retrieval improves oocyte maturation and pregnancy rates. The size of follicles may be an important feature for IVM treatment. The combination of natural-cycle IVF with immature oocyte retrieval followed by IVM is an attractive treatment for young women with all types of infertility without recourse to ovarian stimulation with an acceptable pregnancy rate.

Fertility after breast cancer treatment

In many countries of the developed world, there is an increasing trend toward delay in childbearing from 30 to 40 years of age for various reasons. This is unfortunately concordant with an increasing incidence of breast cancer in women who have not yet completed their family. The current choice for premenopausal women with breast cancer is adjuvant therapy which includes cytotoxic chemotherapy, ovarian ablation (by surgery, irradiation, or chemical ovarian suppression), anti-estrogen therapy, or any combination of these. Although the use of adjuvant therapies with cytotoxic drugs can significantly reduce mortality, it raises issues of the long-term toxicity, such as induction of an early menopause and fertility impairment. The risk of infertility is a potential hardship to be faced by the patients following treatment of breast cancer. The offspring of patients who became pregnant after completion of chemotherapy have shown no adverse effects and congenital anomalies from the treatment, but sometimes high rates of abortion (29%) and premature deliveries with low birth weight (40%) have been demonstrated. Therefore, the issue of recent cytotoxic treatment remains controversial and further research is required to define a “safety period” between cessation of treatment and pregnancy. Preservation of fertility in breast cancer survivors of reproductive age has become an important issue regarding the quality of life. Currently, there are several potential options, including all available assisted technologies, such as in vitro fertilization and embryo transfer, in vitro maturation, oocyte and embryo cryopreservation, and cryopreservation of ovarian tissue. Because increased estrogen levels are thought to be potentially risky in breast cancer patients, recently developed ovarian stimulation protocols with the aromatase inhibitor letrozole and tamoxifen appear to provide safe stimulation with endogenous estrogen. Embryo cryopreservation seems to be the most established fertility preservation strategy, providing a 25–35% chance of pregnancy. In addition, oocyte freezing can be considered as an alternative in patients who are single and in those who do not wish a sperm donor. Although ovarian tissue harvesting appears to be safe, experience regarding ovarian transplantation is still limited due to low utilization, so the true value of this procedure remains to be determined. Nevertheless, in clinical situations in which chemotherapy needs to be started in young patients facing premature ovarian failure, ovarian tissue preservation seems to be a promising option for restoring fertility, especially in conjunction with other options like immature oocyte retrieval, in vitro maturation of oocytes, oocyte vitrification, or embryo cryopreservation. It seems that in vitro maturation is a useful strategy because it improves oocyte or cryopreservation outcome in breast cancer patients undergoing ovarian stimulation for fertility preservation.

In vitro maturation in subfertile women with polycystic ovarian syndrome undergoing assisted reproduction

Polycystic ovarian syndrome (PCOS) occurs in 4% to 7% of all women of reproductive age and 50% of women presenting with subfertility. Subfertility affects 15% to 20% of couples trying to conceive. A significant proportion of these women ultimately need assisted reproductive technology (ART). In vitro fertilisation (IVF) is one of the assisted reproduction techniques employed to raise the chances of achieving a pregnancy. For the standard IVF technique, stimulating follicle development and growth before oocyte retrieval is essential, for which a large number of different methods combining gonadotrophins with a gonadotrophin-releasing hormone (GnRH) agonist or antagonist are used. In women with PCOS, the supra-physiological doses of gonadotrophins used for controlled ovarian hyperstimulation (COH) often result in an exaggerated ovarian response, characterised by the development of a large cohort of follicles of uneven quality, retrieval of immature oocytes, and increased risk of ovarian hyperstimulation syndrome. A potentially effective intervention for women with PCOS-related subfertility involves earlier retrieval of immature oocytes at the germinal-vesicle stage followed by in vitro maturation (IVM). So far, the only data available have derived from observational studies and non-randomised clinical trials. To compare outcomes associated with in vitro maturation (IVM) followed by vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) versus conventional IVF or ICSI, among women with polycystic ovarian syndrome (PCOS) undergoing assisted reproductive technologies (ART). We searched the Menstrual Disorders and Subfertility Group (MDSG) Specialised Register of controlled trials to May 2013 for any relevant trials identified from the title, abstract, or keyword sections. This was followed by a search of the electronic database MEDLINE, EMBASE, LILACS and CINAHL, without language restriction. We also performed a manual search of the references of all retrieved articles; sought unpublished papers and abstracts submitted to international conferences, searched the clinicaltrials.gov and WHO portal registries for submitted protocols of clinical trials, and contacted experts. In addition, we examined the National Institute of Clinical Excellence (NICE) fertility assessment and treatment guidelines and handsearched reference lists of relevant articles (from 1970 to May 2013). All randomised trials (RCTs) on the intention to perform IVM before IVF or ICSI compared with conventional IVF or ICSI for subfertile women with PCOS. Three review authors (CS, MK and NV) independently assessed eligibility and quality of trials. Primary outcome measure was live birth rate per randomised woman. There were no RCTs suitable for inclusion in the review, although there are currently three ongoing trials that have not yet reported results. Though promising data on the IVM technique have been published, unfortunately there is still no evidence from RCTs upon which to base any practice recommendations regarding IVM before IVF or ICSI for women with PCOS. Meanwhile, the results of the above-mentioned ongoing trials are awaited and, of course, further evidence from good quality trials in the field is eagerly anticipated.

A “freeze-all” embryo strategy after in vitro maturation: a novel approach in women with polycystic ovary syndrome?

To assess the efficiency of a "freeze-all" embryo strategy after immature oocyte retrieval (OR) and in vitro maturation (IVM) in patients with polycystic ovary syndrome (PCOS). Retrospective case series. University-based tertiary referral center. Seventy-nine consecutive PCOS patients undergoing IVM followed by vitrified-warmed embryo transfer (ET) over a 2-year period. All patients received 150 IU/d highly purified hMG for 3 consecutive days. There was no hCG trigger given before OR. All day-3 embryos of good morphologic quality were vitrified. Single or double ET was performed in 114 consecutive artificial cycles. The cumulative live birth rate (LBR) per patient was calculated, as well as the projected cumulative LBR. LBR per patient and per retrieved immature oocyte. Mean age, body mass index, and antimüllerian hormone were 28.5 ± 3.5 years, 27.8 ± 7.1 kg/m(2), and 10.3 ± 5.5 μg/L, respectively. In total, 1,526 cumulus-oocyte complexes were retrieved. IVM yielded 800 metaphase II oocytes (52.4%), and 291 day-3 embryos were cryopreserved. Of these, 224 (76.9%) embryos were warmed. One hundred seventy-one survived (76.3%), and 105 ETs were performed. LBR per ET was 16.2% and the cumulative LBR per patient was 21.8%. LBR per retrieved immature oocyte was 1.1%. The projected LBR per patient was 24.2%. IVM followed by a "freeze-all" embryo strategy is a novel approach for women with PCOS. Patients who undergo IVM should be advised that each immature oocyte retrieved yields a 1.1% chance to achieve a live birth.

Overnight maturation of a metaphase I oocyte retrieved from a natural cycle using human tubular fluid: a case report

Since the first successful pregnancy and live birth was achieved through natural cycle in vitro fertilization (IVF) in 1978 [1], there have been many technological advancements and refinements of protocols and techniques through the years [2]. Ovarian hyperstimulation was favored over unstimulated cycles, due to the increased number of embryos available for transfer and hence increased success rate. However, stimulated IVF is also associated with side effects such as ovarian hyperstimulation syndrome [3] and it is costly [4]. For patients who have failed to conceive after repeated stimulated IVF cycles, natural cycle IVF may be a cost-saving alternative [5, 6] which allows patients to avoid further exposure to the side effects associated with stimulated IVF cycles [7, 8]. However, one of the drawbacks of pure natural cycle IVF is its higher cancellations rates (28.9 % per cycle) and lower pregnancy rate (0 to 12.5 % per cycle) [5]. The main reasons for cancellation are premature ovulation before oocyte retrieval, failure to retrieve oocytes and retrieval of immature oocytes. The stage of maturation of the oocyte cannot be determined before retrieval as well. When an immature oocyte is retrieved, it is usually discarded and the cycle is cancelled. To maximize the utilization of the immature oocytes collected from unstimulated cycles, our centre evaluated our protocol and started incubating the immature oocytes overnight in human tubular fluid (HTF) (Irvine Scientific, USA) instead of discarding them. Standard IVF culture media has been shown to be as efficacious as specialized IVM media in ‘rescuing’ immature oocytes retrieved from standard stimulated IVF cycles [9]. When fully matured, the oocytes are fertilized, allowed to develop into embryos in culture and eventually transferred. We report the first case of a successful implantation and pregnancy achieved from this ‘rescue’ protocol in a natural IVF cycle.

A comparison of in-vitro maturation versus antagonist protocols for in-vitro fertilization in women with polycystic ovarian syndrome

Polycystic ovarian syndrome (PCOS) patients are at increased risk of ovarian hyperstimulation syndrome (OHSS) in controlled ovarian hyperstimulation cycles. Inducing oocyte maturation with Gonadotropin Releasing Hormone (GnRH) agonists, in patients stimulated with GnRH antagonist protocols can prevent this complication. In these protocols, ovarian stimulation requires large amounts of gonadotropins. Earlier retrieval of immature oocytes followed by in vitro maturation (IVM) may be a potential alternative intervention for these patients, without exposure to gonadotropins and the risk of OHSS.

Spontaneous pregnancies afterin vitromaturation treatment in two polycystic ovary syndrome patients

Women with polycystic ovary syndrome who need IVF are at increased risk for ovarian hyperstimulation syndrome. Immature oocyte retrieval, and subsequent oocyte in vitro maturation (IVM), is an alternative that does not bear such a risk. We report two cases of spontaneous conception in women with polycystic ovary syndrome-related anovulation, who did not conceive after an IVM treatment. In both patients, spontaneous ovulation and conception occurred within 4 months after the IVM treatment. We hypothesize that this is a beneficial side effect of the ovarian puncture, comparable to the effect of ovarian drilling.

Comparison of assisted reproductive technology outcomes in infertile women with polycystic ovary syndrome:In vitromaturation, GnRH agonist, and GnRH antagonist cycles

ObjectiveWe compared the assisted reproductive technology (ART) outcomes among infertile women with polycystic ovary syndrome (PCOS) treated with IVM, conventional IVF, GnRH agonist, and GnRH antagonist cycles.MethodsThe prospective study included a total of 67 cycles in 61 infertile women with PCOS. The women with PCOS were randomized into three IVF protocols: IVM/IVF with FSH and hCG priming with immature oocyte retrieval 38 hours later (group A, 14 cycles), GnRH agonist long protocol (group B, 14 cycles), and GnRH antagonist multi-dose flexible protocol (group C, 39 cycles). IVF outcomes, such as clinical pregnancy rate (CPR), implantation rate (IR), miscarriage rate (MR), and live birth rate (LBR), were compared among the three groups.ResultsAge, BMI, and basal FSH and LH levels did not differ among the three groups. The number of retrieved oocytes and 2 pronucleus embryos was significantly lower in group A compared with groups B and C. The CPR, IR, MR, and LBR per embryo transfer showed no differences among the three groups. There was no incidence of ovarian hyperstimulation syndrome in group A.ConclusionThe IR, MR, and LBR in the IVM cycles were comparable to those of the GnRH agonist and GnRH antagonist cycles. The IVM protocol, FSH and hCG priming with oocyte retrieval 38 hours later, is an effective ART option that is comparable with conventional IVF for infertile women with PCOS.

Anti-mullerian hormone as a predictive marker for the selection of women for oocyte in vitro maturation treatment

In oocyte in-vitro maturation (IVM) treatments, the chances to achieve a pregnancy are critically dependent on the retrieval of a suitable number of oocytes. In this study, we assessed the ability of circulating levels of anti-mullerian hormone (AMH) to identify normo-ovulatory women suitable for IVM treatment on the basis of the number of retrieved oocytes. Serum AMH was quantified in normo-ovulatory women younger than 39years undergoing IVM treatment. After immature oocyte retrieval and IVM, maximum 3 mature oocytes were used for treatment and all resulting embryos were transferred, as established by law. From 177 cycles, 991 oocytes were recovered. Following IVM, 484 mature oocytes were obtained (50.1%). The overall pregnancy rate per embryo transfer was 16.6% (25/151) and the implantation rate was 10.9% (30/278). Linear regression and receiver operating characteristic (ROC) analyses were applied to identify independent variables and quantify a cut-off AMH value able to identify patients suitable for IVM treatment. An AMH value of 1.28ng/ml was identified as a threshold for the prediction of the retrieval of at least 5 oocytes, with a sensitivity of 93.4% and a specificity of 33.8%. Positive and negative predictive values were 67.6% and 75.0%, respectively. AMH can be adopted to identify women candidate for an IVM treatment from whom a suitable number of oocytes may be retrieved. This is of crucial significance during a non-stimulated cycle, in order to prevent an insufficient oocyte collection and rescue the treatment by implementing a conventional controlled ovarian stimulation.

Health of IVM children

In vitro maturation (IVM) is an emerging type of assisted reproductive technology (ART), in which immature as opposed to mature oocytes are collected from the ovaries and allowed to mature in vitro prior to fertilisation by either standard in vitro fertilisation (IVF) on intracytoplasmic sperm injection (ICSI). Since the first report of a human birth resulting from IVM in 1991 [1], it is estimated that over 1,300 babies have been born following the technique [2] (in contrast to an estimated 3 million babies conceived by standard IVF and ICSI over the last 30 years). IVM offers a number of potential benefits when compared to standard IVF, principally related to the avoidance of high doses of hormonal stimulation with gonadotrophins and GnRH analogues. In patients with polycystic ovary syndrome (PCOS) this avoids the risk of ovarian hyperstimulation syndrome (OHSS). The high costs of hormonal therapy also make IVM cheaper than conventional alternatives. Immature oocyte retrieval, without the need to wait for hormonal stimulation and in vivo maturation, offers the possibility of expedient oocyte cryopreservation for patients diagnosed with cancers without delaying cancer treatment. However, despite recent improvements, pregnancy rates with IVM remain lower than with conventional IVF, and higher miscarriage rates have been reported with IVM [2]. For these reasons IVM has yet to become widely used in the field of ART. Given that IVM is not yet a mainstream ART technique, it is not surprising that there is very limited follow-up data regarding the health of children conceived following its use. This is in contrast to a significant volume of high quality studies that have followed-up children conceived following standard IVF and ICSI. In this article we discuss hypothetical concerns regarding the health of IVM children, and review the few studies published in this area. Health of IVF & ICSI children High multiple pregnancy rates continue to be the single greatest factor adversely impacting upon the outcome of ART pregnancies [3, 4], with resulting increased risk of preterm delivery, low birth weight, and perinatal mortality. However, evidence regarding the health of ART singletons when compared to spontaneously conceived singletons is generally reassuring. The majority of studies investigating the risk of congenital malformations, neurodevelopmental outcome, physical health, growth, and psychosocial well-being in ART children are reassuring [5, 6]. Concerns have been raised that ART children may exhibit higher rates of cancer [7] and rare imprinting disorders [8] (such as Beckwith-Wiedemann Syndrome and Angelman Syndrome), however larger studies are required to confirm these associations. Risks of infertility and cardiovascular disease in later life remain unexplored areas of concern and require follow-up of cohorts of ART children to continue into adulthood.

Effect of incubation with different concentrations and durations of FSH for in-vitro maturation of murine oocytes

The purpose of the study was to evaluate whether a lower concentration of FSH or 2-h incubation with FSH would improve the outcome of in-vitro maturation of oocytes. The immature oocytes were obtained from FVB mice, and were allocated to four groups and incubated in the maturation media for 24 h. The maturation media were supplemented with 10 mIU/ml FSH for 24 h (group 1), 10 mIU/ml FSH for 2 h (group 2), 75 mIU/ml FSH for 24 h (group 3) or 75 mIU/ml FSH for 2 h (group 4). In each group, half of the in-vitro-matured oocytes were fertilized and cultured to blastocysts and the remaining matured oocytes were analysed for growth differentiation factor (GDF)-9 and bone morphogenetic protein (BMP)-15 mRNA to assess the oocyte quality. The maturation rates and oocyte BMP-15 mRNA concentrations were similar among the four groups. The GDF-9 mRNA concentrations were similar in group 2 and group 4. The fertilization and blastocyst rates were higher in groups 2 and 4 than in groups 1 and 3. It is concluded that 2-h incubation with FSH is better than 24-h incubation in terms of the fertilization rate and blastocyst development. Immature oocyte retrieval followed by in-vitro maturation (IVM) is a feasible alternative to IVF. However, the implantation rate of IVM remains lower than that of traditional IVF. The purpose of the study was to evaluate whether a lower concentration of FSH or 2-h incubation with FSH would improve the outcome of IVM. The Immature oocytes were obtained from mice, and were allocated to four groups and incubated in the maturation media for 24 h. The maturation media were supplemented with 10 mIU/ml FSH for 24 h (group 1), 10 mIU/ml FSH for 2 h (group 2), 75 mIU/ml FSH for 24 h (group 3) or 75 mIU/ml FSH for 2 h (group 4). In each group, half of the in-vitro-matured oocytes were fertilized and cultured to blastocysts and the remaining matured oocytes were analysed for growth differentiation factor (GDF)-9 and bone morphogenetic protein (BMP)-15 mRNA to assess the oocyte quality. The maturation rates and oocyte BMP-15 mRNA concentrations were similar between the four groups. The GDF-9 mRNA concentrations were similar in group 2 and group 4. The fertilization and blastocyst rates were higher in groups 2 and 4 than in groups 1 and 3. We conclude that 2-h incubation with FSH is better than 24-h incubation in terms of the fertilization rate and blastocyst development.

Vitrification at the germinal vesicle stage does not affect the methylation profile of H19 and KCNQ1OT1 imprinting centers in human oocytes subsequently matured in vitro

To evaluate the integrity of genomic imprinting in oocytes vitrified at the germinal vesicle (GV) stage and in vitro matured (IVM) after thawing. Clinical research and application. University-based fertility center. Immature oocytes were donated for research by patients who were included in an intracytoplasmic sperm injection program. Immature oocyte retrieval after ovarian stimulation, followed by oocyte vitrification, thawing, and IVM. Methylation profile of H19 and KCNQ1OT1 imprinting control regions, H19DMR and KvDMR1, respectively. Among 184 vitrified GV oocytes, 102 survived thawing (55.4%), 77 (75.5%) of which reached the meiosis II (MII) stage after IVM. One hundred twenty control GV oocytes were only subjected to IVM; 70.8% reached the MII stage. GV vitrified as well as control oocytes acquired full imprint at KvDMR1 after IVM and generally retained the unmethylated state of H19DMR. For the first time, we show that oocyte vitrification does not affect the methylation profile of H19DMR and KvDMR1: during their IVM, vitrified GV oocytes acquire DNA methylation in the maternally imprinted KCNQ1OT1 gene with the same efficiency as fresh GV oocytes; the vitrification process does not alter the unmethylated state of the paternally imprinted H19 gene.