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Successful birth after transfer of blastocysts derived from oocytes of unstimulated woman with regular menstrual cycle after IVM approach.

PURPOSE: To report a delivery after transfer of blastocysts derived from eggs collected following in vivo HCG priming in a patient with regular menstrual cycles undergoing in vitro maturation (IVM) program. METHODS: A woman had regular menstrual cycle and had experience of ovarian hyperstimulation syndrome (OHSS) during a previous conventional IVF-ET cycle. The patient was primed with 10,000 IU HCG 36 h before egg retrieval. After oocyte collection, the maturity of oocytes was evaluated and immature oocytes were cultured in IVM medium. The matured oocytes were fertilized with husband sperm, and normal fertilized eggs were cultured to blastocysts stage until embryo transfer in uteri. RESULTS: Three MII-stage and 13 GV-stage oocytes were collected from the patient. Three mature oocytes were fertilized by conventional IVF. All three fertilized oocytes were developed to blastocysts. Immature oocytes were matured in vitro and insemination was carried out by ICSI. Out of eight fertilized zygotes, two developed to blastocyst stage. Transfer of three expanded blastocysts on Day 6 resulted in pregnancy in the patient and one healthy baby was born. CONCLUSIONS: This report provides an approach to treat infertile women with regular menstrual cycle and high risk of OHSS.

Adult↗

Use of metformin for prevention of ovarian hyperstimulation syndrome: a novel approach.

In the present study, which includes 287 participants, metformin has been used by women undergoing IVF/intracytoplasmic sperm injection for more than 5 weeks before and during treatment and during luteal phase. There was no significant difference in number of gonadotrophins used, days of stimulation, number of oocytes retrieved, and number of embryos replaced. There was no significant difference in clinical pregnancy rate between both groups but there was significant reduction in the incidence of ovarian hyperstimulation syndrome (OHSS) in the group taking metformin. Metformin is a safe, cheap drug that can help in prevention of OHSS.

Adult↗

A guanine nucleotide-binding regulatory protein in human sperm mediates acrosomal exocytosis induced by the human zona pellucida.

Guanine nucleotide-binding regulatory proteins play key intermediary roles in regulating zona pellucida-mediated acrosomal exocytosis in mouse and bull sperm. Since human sperm possess a Gi-like protein and undergo the acrosome reaction in response to the human zona pellucida, we investigated whether this G protein plays a regulatory role in this exocytotic process. Zonae pellucidae isolated from eggs that had been inseminated but had shown no signs of fertilization after retrieval for in vitro fertilization and embryo transfer were pooled into groups of greater than or equal to 50 in order to reduce variability in biological responses due to the possible presence of ZP that had undergone modifications associated with the polyspermy block. Acid-solubilized zonae pellucidae were incubated with capacitated sperm, and the sperm then assessed for the acrosome reaction using both the P. sativum agglutinin and chlortetracycline fluorescence assays; both assays gave similar results. Sperm incubated with solubilized zonae pellucidae at a final concentration of 2, 4, or 6 ZP/microliter underwent acrosomal exocytosis to a similar extent as compared with A-23187. Sperm were incubated with 1 microgram/ml pertussis toxin during capacitation to functionally inactivate the Gi-like protein. Pertussis toxin treatment of sperm did not affect sperm motility and the ability of the cells to bind to structurally intact zonae pellucidae. Pertussis toxin, however, completely inhibited the percentage acrosome reactions induced by solubilized zonae pellucidae. By contrast, the A-23187-induced acrosome reaction was insensitive to PT treatment. Pertussis toxin inhibition of the zona pellucida-induced acrosome reaction occurred in a concentration-dependent manner with maximal effects observed at 100 ng/ml PT.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrosome↗

Relationship between sex hormone-binding globulin and pregnancy outcome in women undergoing controlled ovarian hyperstimulation for assisted reproduction.

The purpose of this study was to determine whether the changes of sex hormone-binding globulin (SHBG) affect the pregnancy outcome in women undergoing controlled ovarian hyperstimulation (COH) for assisted reproduction. Forty-five infertile women who were undergoing pituitary desensitization and COH for in vitro fertilization (IVF) with or without intracytoplasmic sperm injection (ICSI) and 19 women with normal menstrual cycles participated in the study. Fasting blood samples of the follicular and luteal phases, including follicular fluid during oocyte retrieval, were obtained for determination of estradiol (E(2)), progesterone (P(4)), testosterone (T), and SHBG concentrations. The SHBG levels increased progressively during the course of COH, but remained constant throughout normal menstrual cycles. A positive correlation existed between E(2) and SHBG levels in both the follicular and luteal phases. The mean plasma SHBG concentration and E(2)/T ratio were significantly higher, while the level of T and the free androgen index were significantly lower, in the luteal phase of women who conceived than in those who did not conceive following COH. The changes of follicular fluid SHBG level and E(2)/T ratio were similar to those in plasma. We concluded, therefore, that increases in SHBG in the follicular and luteal phases may be a reflection of the functional state of ovarian stimulation, and further that such elevations may influence the pregnancy outcome through the modulation of circulating estrogen and androgen balance during down-regulated COH cycles for IVF/ICSI.

Adult↗

Influence of global sperm DNA methylation on IVF results.

BACKGROUND: In cases of male infertility, routine analysis for sperm characteristics is a poor predictive factor for the segmentation rate and embryo development in assisted reproductive technologies. It is assumed that epigenetic factors could have an influence on the embryo's quality. The aim of this work was to determine the relationship between sperm DNA methylation level and fertilization and pregnancy rates according to the assisted reproduction technique performed. METHODS: A prospective study was undertaken. Ejaculates were obtained from men (n = 63) undergoing an assisted reproduction procedure. 5-Methylcytosine was immunostained with a polyclonal antibody and revealed by fluorescein isothiocyanate. The DNA methylation level was then quantified by flow cytometry. RESULTS: Sixty-three conventional IVF cycles were performed, 760 oocytes were retrieved, an average of 8.1 +/- 4.8 embryos was obtained, and 2.4 embryos were transferred. Neither the fertilization rate nor the rate of good quality embryos was correlated with the DNA methylation level (r = -0.1 and r = -0.08 respectively; not significant). When sperm DNA methylation was >555 arbitrary units, the pregnancy rate was 33.3% compared with 8.3% in the lower (<555) group (P<0.05). CONCLUSION: DNA methylation level in human sperm could represent a new approach to study the ability of sperm to lead to pregnancy in an assisted reproduction procedure, especially when sperm samples with normal characteristics are used.

5-Methylcytosine↗

Validation of the hemizona assay in a monkey model: influence of oocyte maturational stages.

The hemizona assay (HZA) was used in the monkey model to investigate sperm binding to the zona pellucida and to evaluate binding patterns according to various stages of oocyte development. Adult cynomolgus monkeys were superovulated with human menopausal gonadotropin. The oocytes were retrieved by laparoscopy, stored in salt solution, then cut into hemizonae by micromanipulation. Metaphase II oocytes showed significantly tighter binding than prophase I oocytes (P less than 0.001). Metaphase I oocytes showed intermediate binding, significantly different from the other groups (P less than 0.01). It is concluded that: (1) This study demonstrates the feasibility of HZA as a test to evaluate sperm/zona interactions in the monkey, using standards reported for human studies. (2) Oocyte meiotic competence seems to be accompanied by an increase in zona-binding properties.

Animals↗

A comparison of the ovarian response to external stimulation between women with right or left ovary participating in programs of ICSI/ET.

INTRODUCTION: Single ovary women compose a group of special interest in assisted reproduction technologies. The aim of the present study was to investigate whether the response to external stimulation and the outcome in women with a single ovary was affected by the location of the ovary. METHODS: Sixty cycles from 24 women, which stimulated with either hMG or rFSH and a GnRH agonist, were retrospectively studied. Twelve women had right and 12 had left ovary. In all cases, the fertilization was performed by intracytoplasmic sperm injection. RESULTS: There was not found any statistical significant difference between the two groups regarding the days of stimulation, the peak estradiol levels, the number of aspirated follicles, the number of retrieved oocytes, the fertilization rate and the number of transferred embryos. The cumulative embryo score (CES) was also similar in both groups. Six pregnancies in the group of the right ovary and seven in the group of left ovary were achieved. CONCLUSION: According to these results, right and left ovary, in single ovary women under external stimulation, are equally active and potent for reproduction.

Adult↗

Sperm capacitation in the porcine oviduct.

In vitro studies suggests that sperm capacitation occurs in the sperm reservoir (SR) of the pig, with spermatozoa progressing towards the ampullary-isthmic junction (AIJ) around ovulation as a consequence of capacitation/hyperactivation. In contrast, in vivo studies are scarce. Consequently, we determined the degree of capacitation in boar spermatozoa that were retrieved from the SR of sows at well-defined periods of spontaneous standing oestrus, namely pre-, peri- and post-ovulation, using flow cytometry of Merocyanine-540/Yo-Pro-1-loaded spermatozoa. SR-spermatozoa retrieved and incubated in non-capacitating medium (bicarbonate-free mBO [mBO-]) were largely viable (70-85%) and uncapacitated (69-73%), irrespective of the stage of oestrus considered. Those undergoing capacitation were a minor proportion (1-5%) during pre- and peri-ovulation, but they significantly increased (14%) in post-ovulation oestrus. To clarify whether these SR-spermatozoa were able to undergo capacitation under stimuli, sperm aliquots were challenged in vitro either by incubation in a bicarbonate-rich medium (capacitation medium, mBO+), then further in mBO+ with 20% (v/v) of in vivo collected homologous pre-ovulatory isthmic oviductal fluid (IOF), or incubation with hyaluronan (HA, 500 microg/ml). Exposure to mBO+ significantly increased the sub-population of capacitated spermatozoa from the pre- and peri-ovulation SR, indicating that the uncapacitated SR-spermatozoa were responsive to the effector/inducer bicarbonate at levels recorded in peri-ovulatory AIJ/ampulla in vivo. While addition of IOF or HA to SR-spermatozoa incubated in capacitating medium (mBO+) maintained sperm viability without obviously inducing capacitation in pre- or peri-ovulatory SR-spermatozoa, they significantly increased these percentages during post-ovulation, when compared to baseline values of control incubations (mBO-). The results suggest that massive sperm capacitation does not occur in vivo in the porcine SR under spontaneous standing oestrus, particularly during pre- and peri-ovulation, unless spermatozoa are exposed to the effector bicarbonate. Exposure to IOF (and its component HA) under the present experimental conditions, reversed bicarbonate influence during pre- and peri-ovulation and further increased capacitation in post-ovulation, calling for an active role of the intratubal fluid. Furthermore, HA appears to have an active role in the functionality of the SR.

Animals↗

A novel approach for patients at risk for ovarian hyperstimulation syndrome: elective transfer of a single zona-free blastocyst on day 5.

In this preliminary prospective randomized study of 420 patients undergoing ovarian stimulation for IVF/intracytoplasmic sperm injection (ICSI), 17 patients (4%) developed moderate to severe ovarian hyperstimulation syndrome (OHSS). Re-evaluation for OHSS on day 4 and 6 after oocyte retrieval identified one patient with continuous risk for severe OHSS, resulting in cancellation of the transfer (1/17, 5.8%). Prospectively, two of three patients had the zona pellucida of the blastocyst removed by pronase exposure prior to transfer. Significantly more patients became pregnant when a zona-free blastocyst was transferred in comparison to transfer of a single zona-intact embryo (9/11 or 82% versus 1/5 or 20%; P < 0.01). Higher ongoing singleton pregnancy rates were obtained when the zona pellucida was removed prior to the transfer (6/11 and 1/5 respectively). This preliminary prospective randomized study indicates that by prolonging the evaluation time for patients at risk of developing OHSS for up to 6 days after the oocyte retrieval, those patients at risk for developing severe OHSS can be identified. Transferring a single zona-free day 5 embryo (blastocyst) and freezing of the supernumerary embryos offers the patient with moderate OHSS an optimal chance for a singleton pregnancy, while avoiding the serious maternal complications of ovarian hyperstimulation syndrome.

Adult↗

Influence of oocyte preincubation time on fertilization after intracytoplasmic sperm injection.

During the intracytoplasmic sperm injection (ICSI) procedure, the collected oocytes are incubated until just before ICSI. The ideal preincubation time of oocytes was investigated in 544 treatment cycles. Oocyte retrieval was carried out 35 h after human chorionic gonadotrophin administration. Oocytes were cultured for between 1 and 11 h before ICSI. Embryo transfer was performed 48 h after oocyte collection. The survival, fertilization and cleavage rates of injected oocytes indicated no statistically significant differences between oocytes preincubated for different lengths of time. The proportion of good-quality embryos (grades 1 and 2) was lower at 9-11 h of preincubation time than for all the other preincubation times (P < 0.001). No statistically significant differences were detected in the pregnancy rate between each group (mean: 15.9%), although the pregnancy rate at 9-11 h of preincubation time appeared to be low (7.7%). These results suggest that the oocyte retained sufficient potential for fertilization between 1 and 9 h after oocyte collection in ICSI. For the researchers who practise more complex ICSI procedures than IVF, it would be convenient to be able to perform ICSI at any time between 1 and 9 h after oocyte collection.

Adult↗

A prospective, randomized, double-blind study to compare two doses of recombinant human chorionic gonadotropin in inducing final oocyte maturity and the hormonal profile during the luteal phase.

CONTEXT: Different doses of human chorionic gonadotropin (hCG) have been used in various in vitro fertilization (IVF) treatment protocols to achieve final oocyte maturation. There is as yet no agreement on the optimum dose required. OBJECTIVES: The objective of this study was to compare the effectiveness of 250 and 500 microg recombinant hCG (r-hCG), which represented the lower and upper limits of the dose range, in inducing final oocyte maturation during IVF and intracytoplasmic sperm injection cycles. DESIGN: This was a prospective, randomized, double-blind study. SETTING: This study was performed at an IVF clinic in a teaching hospital. PATIENTS: Sixty patients with an indication for intracytoplasmic sperm injection were studied. INTERVENTION: The treatment dose used was 250 or 500 microg r-hCG. MAIN OUTCOME MEASURES: The percentage of metaphase II oocytes retrieved per patient, as an indicator of oocyte maturation, and the hormone profiles of the treatment cycle starting from the day of hCG up to hCG+10 d were the main outcome measures. RESULTS: The percentage of metaphase II oocytes was similar in the two groups (89.3% vs. 86.0%; P = 0.326) despite higher serum and follicular fluid hCG levels on hCG+2 and hCG+4 d, as was the follicular fluid to serum hCG ratio in the 500-microg r-hCG group. Serum estradiol and progesterone levels were comparable initially, but became significantly higher in the 500-microg r-hCG group on hCG+10 d. CONCLUSION: The two doses of r-hCG were equally effective in inducing final oocyte maturation. It remains unclear whether the higher midluteal estradiol and progesterone levels in the 500-microg r-hCG group confer any benefit.

Adult↗

Principal findings from a multicenter trial investigating the safety of follicular-fluid meiosis-activating sterol for in vitro maturation of human cumulus-enclosed oocytes.

OBJECTIVE: To evaluate the safety of applying follicular-fluid meiosis-activating sterol (FF-MAS) in vitro to immature human oocytes. DESIGN: Phase I bicenter, randomized, parallel-group, controlled, partially blinded trial. SETTING: Third-level referral academic centers, including reproductive biology and genetics laboratories. PATIENTS: Endocrinologically normal women with a medical indication for IVF or intracytoplasmic sperm injection, or healthy volunteers. INTERVENTION(S): Subjects were randomized at a ratio 1 to 6 into either conventional GnRH-agonist and recombinant FSH stimulation (IVO) for oocyte retrieval, or minimally stimulated in vitro maturation (IVM) with the use of recombinant FSH. Retrieved immature oocyte cumulus complexes were cultured for 30 or 36 hours in one of six IVM culture conditions containing FF-MAS (range, 0.1-20 microM). Polar body-extruded oocytes from the IVO and IVM groups were processed for chromosomal analysis. MAIN OUTCOME MEASURE(S): The primary endpoint was the incidence of metaphase II stage oocytes with numeric chromosomal abnormalities, using full (spectral karyotyping) or partial (fluorescent in situ hybridization with seven probes) karyotyping or Giemsa count. A secondary objective was to document the frequency of metaphase II oocytes after IVM with FF-MAS supplements. RESULT(S): Oocyte cumulus complexes obtained from the IVO (mean, 8.9) and IVM (mean, 6.2) groups had equal maturation rates. Compared to IVO, exposure of germinal-vesicle oocytes for a maturation period of 30 hours did not increase aneuploidy. An exposure period of 36 hours doubled the aneuploidy rate, but this was significant only for the 20-muM dose of FF-MAS. CONCLUSION: Inclusion of 1-10 microM FF-MAS in a 30-hour IVM protocol is safe.

Adult↗

Effects of caffeine and dbcAMP on zona pellucida penetration by epididymal spermatozoa of cynomolgus monkeys (Macaca fascicularis).

Spermatozoa mature during epididymal transit, acquiring the abilities to swim progressively, fertilize oocytes, and produce viable offspring. In this study, we investigate the capacity of spermatozoa retrieved from the midcorpus and distal cauda regions of the epididymis of the cynomolgus monkey to penetrate homologous zona pellucida. Successful in vitro fertilization by ejaculated macaque sperm is dependent upon the addition of caffeine and dbcAMP. Therefore, the effect of these cyclic nucleotide mediators was also examined in this study. Results of sperm motion analysis indicate no difference in baseline values (without stimulators) for any motion parameter. With the addition of caffeine and dbcAMP, curvilinear velocity significantly increased only for the distal cauda sperm (P = 0.05). Amplitude of the lateral head displacement was significantly increased for distal cauda sperm (P < 0.01); although elevated above baseline, the increase observed after activation by corpus sperm was significantly lower than that achieved by cauda sperm (P < 0.05). The addition of caffeine and dbcAMP was an absolute requirement for zona penetration by both midcorpus and distal cauda sperm. With activation, zona penetration was significantly decreased for corpus sperm compared to cauda sperm (P < 0.001). These results suggest that cynomolgus monkey sperm reaching the midcorpus region of the epididymis have not completed all of the maturational changes requisite for successful fertilization; this immaturity is evidenced by decreased sperm motion and by impedance at the level of zona penetration.

Animals↗

T-helper 2 and 3 type immunity to trophoblast in successful in vitro fertilization-embryo transfer.

OBJECTIVE: To determine whether peripheral blood mononuclear cell (PBMC) secretion of T-helper (Th)-1 type cytokines and Th-2 and Th-3 type cytokines in women undergoing in vitro fertilization-embryo transfer (IVF-ET) is associated with therapeutic failure and success, respectively. DESIGN: Cohort study. SETTING: Academic medical center. PATIENT(S): One hundred one women undergoing IVF-ET and 19 fertile controls. INTERVENTION(S): Peripheral blood was obtained from women undergoing IVF-ET before oocyte retrieval and from 19 nonpregnant fertile controls. The PBMCs were cultured in the presence or absence of a protein extract from either a trophoblast cell line or sperm membrane. MAIN OUTCOME MEASURE(S): Supernatants from PBMC cultures were tested by enzyme-linked immunoabsorbent assay (ELISA) for the Th-1 type cytokines tumor necrosis factor-alpha (TNF)-alpha) and interferon-gamma (IFN)-gamma), the Th-2 type cytokines interleukin (IL)-6 and IL-10, and the Th-3 type cytokine transforming growth factor (TGF)-beta1. RESULT(S): Levels of IL-6 and IL-10 were significantly higher in controls than in infertile women with endometriosis, and levels of IL-10 were higher in controls than in women with unexplained infertility. No differences were found in unstimulated levels of TNF-alpha, IFN-gamma, or TGF-beta1 between infertile patients and controls. In trophoblast-stimulated PBMC cultures, levels of TGF-beta1 were significantly lower in subjects who experienced failed compared with ongoing pregnancies. CONCLUSION(S): Baseline PBMC secretion of IL-6 and IL-10 is higher in fertile controls than in women with endometriosis, and IL-10 secretion is also higher than in women with unexplained infertility. Trophoblast-stimulated PBMC secretion of TGF-beta1 is positively associated with the establishment of successful pregnancy in women undergoing IVF-ET. Our study provides novel evidence to support a faciliatory role of Th-2 and Th-3 type responses to trophoblast in early pregnancy.

Analysis of Variance↗

The human genome encodes 10 alpha-crystallin-related small heat shock proteins: HspB1-10.

To obtain an inventory of all human genes that code for alpha-crystallin-related small heat shock proteins (sHsps), the databases available from the public International Human Genome Sequencing Consortium (IHGSC) and the private Celera human genome project were exhaustively searched. Using the human Hsp27 protein sequence as a query in the protein databases, which are derived from the predicted genes in the human genome, 10 sHsp-like proteins were retrieved, including Hsp27 itself. Repeating the search procedure with all 10 proteins and with a variety of more distantly related animal sHsps, no further human sHsps were detected, as was the case when searches were performed at deoxyribonucleic acid level. The 10 retrieved proteins comprised the 9 earlier recognized human sHsps (Hsp27/HspB1, HspB2, HspB3, alphaA-crystallin/HspB4, alphaB-crystallin/HspB5, Hsp20/HspB6, cvHsp/HspB7, H11/HspB8, and HspB9) and a sperm tail protein known since 1993 as outer dense fiber protein 1 (ODF1). Although this latter protein probably serves a structural role and has a high cysteine content (14%), it clearly contains an alpha-crystallin domain that is characteristic for sHsps. ODF1 can as such be designated as HspB10. The expression of all 10 human sHsp genes was confirmed by expressed sequence tag (EST) searches. For Hsp27/HspB1, 2 retropseudogenes were detected. The HspB1-10 genes are dispersed over 9 chromosomes, reflecting their ancient origin. Two of the genes (HspB3 and HspB9) are intronless, and the others have 1 or 2 introns at various positions. The transcripts of several sHsp genes, notably HspB7, display low levels of alternative splicing, as supported by EST evidence, which may result in minor amounts of isoforms at the protein level.

Amino Acid Sequence↗

[A study of aspermia-related genes by genchips and analysis of the RAP1A gene].

OBJECTIVE: To study the differential gene expression profiles between the normal and aspermia human testes by genechips. METHODS: Probes were prepared from mRNA extracted from both normal and aspermia testes and employed on Biostar H-40s genechips to detect the differential gene expression profiles. A distinctly up-regulated gene RAP1A was analyzed by bibliogrphic retrieval. RESULTS: Six hundred and twenty-three differential expressed genes were found, among which the distinctly up-regulated gene RAP1A was closely related to human sperm regulation. CONCLUSIONS: Screening the differential gene expression profiles between the normal and aspermia human testes by genechips can be used in the study of aspermia-related genes.

Adult↗

The origin of spermatozoa does not affect intracytoplasmic sperm injection outcome.

OBJECTIVE: To assess whether the origin of spermatozoa, ejaculate or testicular, affects intracytoplasmic sperm injection (ICSI) outcome. STUDY DESIGN: Retrospective study of 890 consecutive first ICSI and embryo transfer cycles done for male infertility. The ICSI outcome of ejaculated spermatozoa (n=780) and testicular spermatozoa retrieved from patients with obstructive azoospermia (n=43), non-obstructive azoospermia (n=53) and severe oligoasthenoteratozoospermia (n=14) were compared by using chi-square test, independent t-test and ANOVA with Bonferroni test. RESULTS: All azoospermic males had a diagnostic testicular biopsy at least 6 months before the ICSI procedure. Spermatozoa were successfully retrieved in all 43 patients with obstructive azoospermia and in 72.6% of 73 non-obstructive cases. The cycle characteristics of the four groups were similar apart from a younger mean female age in the non-obstructive azoospermia group when compared with the ejaculated spermatozoa group. The fertilization, implantation and clinical pregnancy rates were comparable among the four groups. CONCLUSION: Testicular spermatozoa recovered from patients with obstructive and all types of non-obstructive azoospermia were as much as effective as ejaculated spermatozoa in ICSI.

17 alpha-Hydroxyprogesterone Caproate↗

[The applied value of rescue intracytoplasmic sperm injection after complete fertilization failure during in vitro fertilization cycles].

OBJECTIVE: To discuss the applied value of rescue intracytoplasmic sperm injection(ICSI) after complete fertilization failure during in vitro fertilization (IVF) cycles. METHODS: After 16-18 h co-culture with sperm, all the unfertilized oocytes with the first polar body were re-fertilized by ICSI. RESULTS: After rescue ICSI, the abnormal fertilization rate was 17.9% and the normal fertilization rate was 42.7%. Twenty-four hours later, the normal cleavage rate of the normal fertilized oocytes was 79.6%. On the day of embryo transfer, the good-quality rate of embryos was 29.7% (22/74). A mean number of 3.4 (54/16) embryos were transferred to the patient during each cycle. Clinical pregnancy was found in 3 cases out of the 16 (18.8%). CONCLUSION: The applied value of rescue ICSI may be correlated with the number and maturity of oocytes on the retrieval day.

Female↗