The Effect of Preincubation Period of Oocytes on Nuclear Maturity, Fertilization Rate, Embryo Quality, and Pregnancy Outcome in IVF and ICSI

Purpose : To clarify the effect of preincubation of oocytes on the results of IVF and ICSI. Methods : A total of 176 IVF and 64 ICSI cycles received long protocol ovarian stimulation. The oocytes were incubated for 1–8 h before insemination or sperm injection. Metaphase II (MII) percentage was evaluated in the ICSI arm; fertilization rates, embryo quality, and pregnancy outcomes were analyzed in both IVF and ICSI arms according to the preincubation period duration of oocytes. Results : The MII percentage of the ICSI arm was significantly lower (P < 0.05) in the group with preincubation period of <2.5 h. The fertilization rates in groups with preincubation for 2.5–5.5 h were significantly higher (P < 0.001) for IVF. Embryo quality and pregnancy outcomes were not significantly different between the IVF or ICSI arm. Conclusions : The preincubation of oocytes for at least 2.5 h is beneficial to both IVF and ICSI outcomes by increasing the nuclear maturity of oocytes.     

Effect of gonadotrophin stimulation on mouse oocyte quality and subsequent embryonic development in vitro

In-vivo-matured oocytes were collected from naturally ovulated and superovulated [pregnant mare's serum gonadotrophin (PMSG) + human chorionic gonadotrophin (HCG)] mice. Immature oocytes were retrieved from naturally cycling mice and from mice primed with PMSG. The percentages of cleavage and blastocyst formation were significantly different (P < 0.05) between in-vivo- and in-vitro-matured oocytes. Blastocyst formation rate was significantly higher (P < 0.05) in immature oocytes derived from PMSG-primed mice, and the percentages of oocytes with comet tails, and their length, were significantly higher and longer respectively in in-vitro-matured oocytes. Total cell numbers of blastocysts were also significantly different (P < 0.05) between in-vivo- and in-vitro-matured oocytes, but there were also no differences in ratio of trophectoderm (TE)/inner cell mass (ICM). In conclusion, in-vivo-matured mouse oocytes were more competent than those matured in-vitro, perhaps due to a lesser degree of DNA damage. Embryonic development capacity of in-vivo-matured oocytes is not promoted by ovarian stimulation. Gonadotrophin priming prior to immature mouse oocyte retrieval is beneficial to subsequent embryonic development.     

Are Cumulus Cells Necessary for the Spontaneous Maturation of Germinal Vesicle-Stage Oocytes to Metaphase II

Purpose: In this study we investigated the need of the support from cumulus cells for germinal-vesicle (GV) oocytes collected from stimulated ovaries to complete their maturation to metaphase II (MII). Methods: We compared the maturation rate of GV oocytes after coculture with cumulus cells (study group) with their spontaneous maturation in culture medium alone (control group). Results: Sixty-four and nine-tenths percent of the GV oocytes matured to metaphase II in the coculture group, and of these, 43.5% gave normal 2pn zygotes following intracytoplasmic sperm injection (ICSI), while 73.8% of the GV oocytes spontaneously matured to the MII stage and 30% of these reached the zygote stage after ICSI. Conclusions: It is probable that a follicular factor is responsible for this arrested maturation in the human and that maturation occurs spontaneously when the oocytes are separated from their follicular fluid environment after collection.     

Deliveries of babies with normal health derived from oocytes with smooth endoplasmic reticulum clusters

PurposeTo examine the impact on development of derived embryos from smooth endoplasmic reticulum clusters (SERC) in human metaphase II (MII) oocytes.MethodsRetrospective analysis at Kyono ART Clinic. Comparison of embryological development, pregnancy, live birth and fetal malformation between oocytes with SERC (the SERC(+) group) and those without (the SERC(−) group) in 2,158 patients (3,758 cycles) after ICSI.ResultsFertilization and implantation rate were significantly lower in SERC(+) MII oocytes than in SERC(−) MII oocytes. After the transfer of fresh and vitrified embryos derived from SERC(+) oocytes, 14 pregnancies resulted in 14 healthy babies, including 2 from fresh embryo transfer (ET) and 12 from vitrified-warmed ET, with no malformations.Conclusion(s)The presence of SERC in MII oocytes was associated with significantly lower fertilization rates and implantation rates than seen in SERC(−) MII oocytes within SERC (+) cycles. However, SERC had no impact on post-implantation development as well as neonatal outcome.     

How effective are the non-conventional ovarian stimulation protocols in ART? A systematic review and meta-analysis

PurposeTo compare the effectiveness of starting the ovarian stimulation on the early follicular phase (“Conventional”) with the newer range of non-conventional approaches starting in the luteal phase (“Luteal”), random-start, and studies implementing them in DuoStim (“Conventional”+“Luteal”).MethodsSystematic review. We searched CENTRAL, PubMed, and Embase, on March 2020. We included randomized and non-randomized controlled trials that compared “Luteal,” random-start ovarian stimulation or DuoStim with “Conventional”; we analyzed them by subgroups: oocyte freezing and patients undergoing ART treatments, both, in the general infertile population and among poor responders.ResultsThe following results come from a sensitivity analysis that included only the low/moderate risk of bias studies. When comparing “Luteal” to “Conventional,” clinically relevant differences in MII oocytes were ruled out in all subgroups. We found that “Luteal” probably increases the COH length both, in the general infertile population (OR 2.00 days, 95% CI 0.81 to 3.19, moderate-quality evidence) and in oocyte freezing cycles (MD 0.85 days, 95% CI 0.53 to 1.18, moderate-quality evidence). When analyzing DuoStim among poor responders, we found that it appears to generate a higher number of MII oocytes in comparison with a single “Conventional” (MD 3.35, 95%CI 2.54–4.15, moderate-quality evidence).ConclusionOverall, this systematic review of the available data demonstrates that in poor responders, general infertile population and oocyte freezing for cancer stimulation in the late follicular and luteal phases can be utilized in non-conventional approaches such as random-start and DuoStim cycles, offering similar outcomes to the conventional cycles but potentially with increased flexibility, within a reduced time frame. However, more well-designed trials are required to establish certainty.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10815-020-01966-5.     

High proportion of immature oocytes in a cohort reduces fertilization,embryo development,pregnancy and live birth rates following ICSI

Research questionOoplasmic maturity has been studied for some time, but remains poorly defined. This study aimed to evaluate metaphase II (MII) oocyte competence in terms of fertilization, embryo development and cycle outcomes, according to the oocyte maturity ratio.DesignCouples treated by intracytoplasmic sperm injection (ICSI) between 1993 and 2017 with female partners ≤35 years old were included. Cycles were divided into four groups according to proportion of MII oocytes at the time of retrieval: optimal (76–100%), adequate (51–75%), partial (26–50%) and minimal (1–25%).ResultsA total of 7672 ICSI cycles (optimal: 4838; adequate: 2252; partial: 518; minimal oocyte maturity: 64) were included, in which 95,667 MII oocytes were injected using ejaculated spermatozoa. The decreasing proportion of MII significantly reduced normal fertilization (two pronuclei) (78.9% to 71.3%; P < 0.0001) with a corresponding increase in digynic three-pronuclei that rose from 2.6% in the optimal group to 4.7% in the minimal group (P = 0.003). Implantation (33% to 17%; P < 0.0001), clinical pregnancy (63.6% to 37.5%; P < 0.0001) and live birth rates (49.2% to 26.6%; P < 0.0001) were affected by the decreasing proportion of MII oocytes.ConclusionsA high proportion of immature sibling oocytes in the retrieved cohort affects the fertilization rate and embryo developmental competence of MII inseminated oocytes, clinical pregnancy and live birth rates, suggesting that, in addition to nuclear maturity, ooplasmic and membrane maturity are required for developmental competence of MII oocytes. These findings may provide guidance toward ovarian stimulation protocols aimed at achieving a greater proportion of MII oocytes, leading to higher fertilization rates and better pregnancy outcomes.     

Increased DNA damage and repair deficiency in granulosa cells are associated with ovarian aging in rhesus monkey

PurposeOvarian aging is closely tied to the decline in ovarian follicular reserve and oocyte quality. During the prolonged reproductive lifespan of the female, granulosa cells connected with oocytes play critical roles in maintaining follicle reservoir, oocyte growth and follicular development. We tested whether double-strand breaks (DSBs) and repair in granulosa cells within the follicular reservoir are associated with ovarian aging.MethodsOvaries were sectioned and processed for epi-fluorescence microscopy, confocal microscopy, and immunohistochemistry. DNA damage was revealed by immunstaining of γH2AX foci and telomere damage by γH2AX foci co-localized with telomere associated protein TRF2. DNA repair was indicated by BRCA1 immunofluorescence.ResultsDSBs in granulosa cells increase and DSB repair ability, characterized by BRCA1 foci, decreases with advancing age. γH2AX foci increase in primordial, primary and secondary follicles with advancing age. Likewise, telomere damage increases with advancing age. In contrast, BRCA1 foci in granulosa cells of primordial, primary and secondary follicles decrease with monkey age. BRCA1 positive foci in the oocyte nuclei also decline with maternal age.ConclusionsIncreased DSBs and reduced DNA repair in granulosa cells may contribute to ovarian aging. Discovery of therapeutics that targets these pathways might help maintain follicle reserve and postpone ovarian dysfunction with age.

Electronic supplementary material

The online version of this article (doi:10.1007/s10815-015-0483-5) contains supplementary material, which is available to authorized users.     

Effects of letrozole or tamoxifen coadministered with a standard stimulation protocol on fertility preservation among breast cancer patients

PurposeTo assess the effects of letrozole or tamoxifen coadministration on fertility preservation treatment outcomes.MethodsRetrospective cohort study of 118 breast cancer patients undergoing fertility preservation treatment between 2008 and 2018. Patients who received letrozole (n = 36) or tamoxifen (n = 30) were compared to controls (n = 52) who underwent standard ovarian stimulation protocols. The primary outcome measures included the number of retrieved oocytes, mature oocytes (MII), fertilization, and top-quality embryo rates. The secondary outcome measures included duration of stimulation, gonadotropin dose and peak estradiol level.ResultsThe number of oocytes retrieved, MII oocytes, fertilization rate, duration of stimulation, or gonadotropin dose were similar in the letrozole and tamoxifen groups, compared to controls. Top-quality embryo rate was lower in the tamoxifen group compared to controls (25% vs 39.4%, respectively, P = 0.034). The abnormal fertilization rate was higher in the letrozole group compared to controls (7.8% vs 3.60%, respectively, P = 0.015). A stepwise logistic regression analysis revealed that letrozole and peak estradiol were significantly associated with abnormal fertilization (OR 11.94; 95% CI 2.35–60.4, P = 0.003 for letrozole and OR 1.075; 95% CI 1.024–1.12, P = 0.004 per 100 unit change in estradiol).ConclusionsThere may be a negative effect of letrozole or tamoxifen on fertilization and embryo quality, in fertility preservation cycles. Further studies are needed to confirm these findings.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10815-020-02030-y.     

Relationship of the Human Cumulus-Free Oocyte Maturational Profile with In Vitro Outcome Parameters After Intracytoplasmic Sperm Injection

Purpose: We investigated whether the human oocyte maturational profile at the removal of cumulus/corona cells affects the fertilization rate and subsequent embryo quality after intracytoplasmic sperm injection. Methods: A total of 1011 oocytes from 150 cycles was included in this retrospective analysis. Cumulus-free oocytes that were in prophase or metaphase I of meiosis at the removal of cumulus/corona cells were incubated in vitro until they reached metaphase II (in vitro-matured oocytes) and were then immediately injected with a single spermatozoa. Oocytes that were in metaphase II at the removal of cumulus/corona cells (MII oocytes) received sperm injection after 3–4 hr of preinjection incubation. Results: The fertilization rate of the MII oocytes was significantly higher than that of in vitro-matured oocytes (81 vs 62%; P < 0.001). The cleavage rates were similar in the two groups (MII oocytes, 94%; in vitro-matured oocytes, 91%). However, MII oocytes had significantly higher percentages of good-quality embryos (grade 1–3 embryos, 87 vs 58%, P < 0.001) and embryos with high cumulative embryo scores (score 10–32 embryos, 62 vs 33%, P < 0.001). The mean cumulative embryo score of MII oocytes after fertilization was also higher than that of in vitro-matured oocytes (12.1 ± 3.8 vs 8.8 ± 3.4; P = 0.014). Conclusions: MII oocytes that extruded the first polar body at the removal of cumulus/corona cells had better fertilization rates and embryo morphology than in vitro-matured oocytes that extruded the first polar body following the removal of cumulus/corona cells and in vitro culture.     

DNA repair in primordial follicle oocytes following cisplatin treatment

Purpose

Genotoxic chemotherapy and radiotherapy can cause DNA double stranded breaks (DSBs) in primordial follicle (PMF) oocytes, which then undergo apoptosis. The development of effective new fertility preservation agents has been hampered, in part, by a limited understanding of DNA repair in PMF oocytes. This study investigated the induction of classical DSB repair pathways in the follicles of wild type (WT) and apoptosis-deficient Puma^-/- mice in response to DSBs caused by the chemotherapy agent cisplatin.

Methods

Adult C57BL/6 WT and Puma^-/- mice were injected i.p. with saline or cisplatin (5 mg/kg); ovaries were harvested at 8 or 24 h. Follicles were counted, and H2A histone family member (γH2AX) immunofluorescence used to demonstrate DSBs. DNA repair protein RAD51 homolog 1 (RAD51) and DNA-dependent protein kinase, catalytic subunit (DNA-PKcs) immunofluorescence were used to identify DNA repair pathways utilised.

Results

Puma^-/- mice retained 100% of follicles 24 h after cisplatin treatment. Eight hours post-treatment, γH2AX immunofluorescence showed DSBs across follicular stages in Puma^-/- mice; staining returned to control levels in PMFs within 5 days, suggesting repair of PMF oocytes in this window. RAD51 immunofluorescence eight hours post-cisplatin was positive in damaged cell types in both WT and Puma^-/- mice, demonstrating induction of the homologous recombination pathway. In contrast, DNA-PKcs staining were rarely observed in PMFs, indicating non-homologous end joining plays an insignificant role.

Conclusion

PMF oocytes are able to conduct high-fidelity repair of DNA damage accumulated during chemotherapy. Therefore, apoptosis inhibition presents a viable strategy for fertility preservation in women undergoing treatment.

     

Growth hormone supplementation during ovarian stimulation improves oocyte and embryo outcomes in IVF/PGT-A cycles of women who are not poor responders

PurposeTo determine the effect of human growth hormone (GH) supplementation during ovarian stimulation in women undergoing IVF/PGT-A cycles, who do not meet the Bologna criteria for poor ovarian response (POR).MethodsThis is a retrospective cohort study of 41 women with suboptimal outcomes in their first cycle of IVF/PGT-A including lower than expected number of MII oocytes, poor blastulation rate, and/or lower than expected number of euploid embryos for their age, who underwent a subsequent IVF/PGT-A cycle with the same fixed dose gonadotropin protocol and adjuvant GH treatment. Daily cotreatment with GH started with first gonadotrophin injection. The IVF cycle outcomes were compared between the control and GH cycle using the Wilcoxon-Signed Rank test.ResultsThe total number of biopsied blastocysts (mean ± SD; 2.0 ± 1.6 vs 3.5 ± 3.2, p = 0.009) and euploid embryos (0.8 ± 1.0 vs 2.0 ± 2.8, p = 0.004) were significantly increased in the adjuvant GH cycle compared to the control cycle. The total number of MII oocytes also trended to be higher in the GH cycle (10.2 ± 6.3 vs 12.1 ± 8.3, p = 0.061). The overall blastulation and euploidy rate did not differ between the control and treatment cycle.ConclusionOur study uniquely investigated the use of adjuvant GH in IVF/PGT-A cycles in women without POR and without a priori suspicion for poor outcome based on their clinical parameters. Our study presents preliminary evidence that GH supplementation in these women is beneficial and is associated with an increased number of blastocysts for biopsy and greater number of euploid embryos for transfer.     

Is corifollitropin alfa effective in controlled ovarian stimulation among all poor ovarian responders? A retrospective comparative study

Abstract

Several studies have compared the effectiveness of corifollitropin alfa versus daily gonadotropins in poor ovarian responders (PORs) undergoing controlled ovarian stimulation (COS), showing conflicting results in terms of IVF outcomes. Given the heterogeneity of patients included in the classification of POR according to ‘Bologna criteria’, the aim of this study was to evaluate the impact of corifollitropin alfa in two different categories of POR distinguished according to patients’ antral follicle count (AFC). We retrospectively evaluated 104 infertile POR, split into two groups according to AFC (Group A?≤?5; Group B?>?5) and subgroups according to the ovarian stimulation regimen (corifollitropin alfa plus daily gonadotropins (Subgroup 1) versus daily gonadotropins alone (Subgroup 2)). Outcome measures were total oocytes, MII oocytes, total embryos, follicular output rate (FORT), implantation rate (IR), clinical pregnancy rate (CPR), miscarriage rate (MR), and live birth rate (LBR). Subgroup A1 experienced a lower number of total oocytes, MII oocytes, total embryos, and FORT (p?<?.05) in comparison to Subgroup A2, while no difference was found when comparing Subgroups B1 and B2. No difference was found between subgroups even in terms of IR, CPR, MR, and LBR. In conclusion, corifollitropin alfa may be as effective as daily gonadotropins in POR with AFC?>?5 undergoing COS, while it might be inferior to daily gonadotropins in POR with AFC?≤?5.     

Detection of aneuploidy in human oocytes and corresponding first polar bodies by fluorescent in situ hybridization

Purpose: The purpose of the study was to investigate the reliability of the fluorescent in situ hybridization (FISH) analysis of the first polar body (IPB) for cytogenetic evaluation of human oocytes as a method of choice in preimplantation diagnosis of chromosomal aneuploidies. Design: Human unfertilized oocytes and their extruded IPB were analyzed using the directly labeled fluorescence alpha-satellite DNA probes to chromosomes X and 18. Results: Paired signals for chromosomes X and 18 were observed in the second meiotic prophase (MII) of unfertilized oocytes and their extruded IPB. In the series of 156 unfertilized oocytes in which the number of X chromosome-and chromosome 18-specific signals were analyzed in both MII and IPB, five nondisjunction events have been detected, with corresponding signals in MII and their IPB: missing signals in MII corresponded to extra signals in their IPB and extra signals in MII corresponded to missing signals in IPB. In one oocyte chromosome 18 nondisjunction was detected, with both chromosome 18 signals in MII and no chromosome 18 signal in IPB. In four oocytes chromatid malsegregations for chromosome X or chromosome 18 were detected: in two oocytes, three of four chromosome 18 signals were present in MII, with only one in IPB, and in the other two oocytes, three of four chromosome signals were present in MII, with only one left in IPB. Conclusions: The data suggest the possibility of detecting chromosomal aneuploidy in oocytes through cytogenetic analysis of their corresponding IPB by FISH as a possible approach for preimplantation diagnosis of major chromosomal trisomies.Presented in part at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, Vienna, Austria, April 3–7, 1995.     

Correlation between human follicular diameter and oocyte outcomes in an ICSI program

Purpose: To determine the correlation between the follicular sizes and oocyte recovery, metaphase II oocyte recovery, fertilization rate and good embryo quality from mature and immature oocytes in an intracytoplasmic sperm injection (ICSI) program. Methods: 991 follicles obtained from 72 ICSI cycles were classified into three groups according to their diameters as measured by transvaginal ultrasound including group A (<10 mm), group B (10–14 mm), and group C (>14 mm). All obtained oocytes were classified according to their nuclear maturation: germinal vesicle (GV), metaphase I (MI) and metaphase II (MII). Mature oocytes underwent ICSI while immature oocytes were further cultured until maturity before ICSI was performed. The rates of fertilization and good quality embryos at day 3 were evaluated. Results: A progressive and significant increase in the rates of oocyte recovery and MII oocyte recovery were observed from group A follicles compared to the other groups (p < 0.001). The fertilization rate of mature and in vitro matured oocytes, as well as the rate of good quality embryos showed a tendency to increase from group A to group C follicles, but not significantly. The corresponding fertilization rates were 78 and 55.3% (p < 0.001) for mature and in vitro matured oocytes, respectively. Conclusion: Collection of oocytes from small follicles, especially with a mean diameter less than 10 mm, and in vitro maturation of immature oocytes before fertilization may allow the total number of good quality and transferable embryos to be increased.     

Aging attenuates the ovarian circadian rhythm

ObjectiveTo study the effect of aging on ovarian circadian rhythm.DesignHuman and animal study.SettingUniversity hospital and research laboratory.Patients/animalsHuman granulosa cells were obtained by follicular aspiration from women undergoing in vitro fertilization (IVF), and ovarian and liver tissues were obtained from female C57BL/6 mice.Intervention(s)None.Main outcome measure(s)Expression of circadian genes in young and older human granulosa cells and circadian rhythm in ovaries and livers of young and older mice.Result(s)All examined circadian clock genes in human granulosa cells showed a downward trend in expression with aging, and their mRNA expression levels were negatively correlated with age (P < 0.05). Older patients (≥ 40 years of age) had significantly reduced serum anti-Müllerian hormone (AMH) levels. Except for Rev-erbα, all other examined circadian clock genes were positively correlated with the level of AMH (P < 0.05). The circadian rhythm in the ovaries of older mice (8 months) was changed significantly relative to that in ovaries of young mice (12 weeks), although the circadian rhythm in the livers of older mice was basically consistent with that of young mice.Conclusion(s)Lower ovarian reserve in older women is partially due to ovarian circadian dysrhythmia as a result of aging.Electronic supplementary materialThe online version of this article (10.1007/s10815-020-01943-y) contains supplementary material, which is available to authorized users.     

Effects of high-dose and multiple-dose gonadotropin stimulation on mouse oocyte quality as assessed by preimplantation development following in vitro fertilization

The effects of increasing the level of ovarian stimulation on preimplantation embryonic development were assessed using a mouse in vitro fertilization system. When F₁ hybrid (C57BL/6 × CBA/Ca) mice received a single injection of 5 IU pregnant mare's serum gonadotropin (PMSG) followed 60 hr later by 5 IU human chorionic gonadotropin (hCG) approximately 50% of the resultant postovulatory oocytes developed to the blastocyst stage following in vitro fertilization. Increasing the single dose of PMSG to 10 or 15 IU resulted in significant reductions in the frequency of development to the blastocyst stage. When one or two additional doses of 5 IU PMSG were administered 24 and 48 hr after an initial injection of 5 IU, lower frequencies of oocytes with the potential for full preimplantation development were again observed. This reduction in gamete quality was significantly greater when the final dose of PMSG was administered only 12 hr prior to hCG. The results suggest that excessive gonadotropin stimulation may compromise the quality of the preimplantation embryos obtained following in vitro fertilization and that the timing of gonadotropin administration may also be critical.     

Anti-mullerian hormone level is a reliable predictor for cycle cancellation in ICSI

ObjectivesTo evaluate serum AMH level as a predictor of ovarian response to stimulation and outcome of ICSI.Design of the studyA prospective clinical study.Setting and timeMansoura general hospital and Benha university hospital, during the period from October 2007 to May 2008.Patients and methodsSixty women aged 20–39years scheduled for ICSI were included in the study. Cases of polycystic ovaries were excluded. Serum AMH level was determined using an enzyme-amplified two-site immunoassay. The stimulation protocol used was the short protocol.Outcome measuresCorrelation between the AMH level and the number of retrieved MII oocytes; the AMH level in cancelled and completed cycles and in non-pregnant and pregnant and its value in prediction of these variables.ResultsThere was a highly significant positive correlation between AMH level and number of retrieved MII oocytes (R²=0.33, P<0.001). Levels were highly significant lower in cancelled than in completed cycles (P=0.0143), and non-significant lower in non-pregnant than in pregnant (P=0.0519). The AMH level was a fair test for discrimination between cancelled and completed cycles (ROC–AUC 0.747, 95% CI 0.618–0.850), but it was a poor test for discrimination between non-pregnant and pregnant (ROC–AUC 0.659, 95% CI 0.526–0.777). Cutoff level AMH of 0.9ng/ml had the best sensitivity (80%) and specificity (64%) in predicting poor response and cycle cancellation (P<0.01), but it was of no value in predicting non-pregnancy.ConclusionsAMH is a reliable marker for ovarian response to stimulation as regards the number of retrieved MII oocytes and cancelled or completed cycles, but not for the success of ICSI as regards non-pregnancy or pregnancy.     

Pregnancy and delivery after in vitro maturation of naked ICSI GV oocytes with GH and transfer of a frozen thawed blastocyst: case report

Purpose : To determine if GV oocytes, collected at the time of ICSI, can be matured in vitro and rescued for therapeutic treatment. A patient for whom all the collected oocytes at the GV stage after a classical COH protocol were matured in vitro with GH. Method : All the naked oocytes were matured in a culture medium (ISM2) containing 15% patient serum +1.6 units of GH (Saizen) per millilitre. Oocytes were incubated overnight at 37°C. The MII oocytes obtained were micro-injected. A fresh transfer was performed and a supernumerary blastocyst was frozen. Results : The patient was pregnant and delivered a healthy girl after transfer of the frozen/thawed blastocyst. The baby girl is now 2 years old. Conclusion : In vitro maturation with GH allows rescuing naked GV oocytes collected at the time of ICSI. GH action does not pass through the cumulus cells. According to the possible lack of synchrony between the embryo and the uterus, we recommend to freeze the embryos obtained and to replace them in a controlled cycle.