Intracytoplasmic Sperm Injection Failure Research Articles

Use of testicular sperm in couples with SCSA-defined high sperm DNA fragmentation and failed intracytoplasmic sperm injection using ejaculated sperm.

Sperm DNA fragmentation (SDF) has been linked with male infertility, and previous studies suggest that SDF can have negative influence on pregnancy outcomes with assisted reproduction. We performed a retrospective review of consecutive couples with a high SDF level that had intracytoplasmic sperm injection (ICSI) using testicular sperm (T-ICSI). We compared the T-ICSI outcomes to that of two control groups: 87 couples with failed first ICSI cycle and who had a second ICSI cycle using ejaculated sperm (Ej-ICSI), and 48 consecutive couples with high sperm chromatin structure assay (SCSA)-defined SDF (>15%) that underwent an ICSI cycle using ejaculated sperm after one or more failed ICSI cycles (Ej-ICSI-high SDF). The mean number of oocytes that were retrieved and the total number of embryos were not different among the three groups. The mean number of transferred embryos in the T-ICSI group was higher than the Ej-ICSI group but not significantly different than the Ej-ICSI-high SDF group (1.4, 1.2, and 1.3, respectively, P < 0.05). Clinical pregnancy rate in the T-ICSI group was not significantly different than the Ej-ICSI and Ej-ICSI-high SDF groups (48.6%, 48.2%, and 38.7%, respectively, P > 0.05). No significant difference was found in live birth rate when comparing T-ICSI to Ej-ICSI and Ej-ICSI-high SDF groups. The results suggest that pregnancy outcomes and live birth rates with T-ICSI are not significantly superior to Ej-ICSI in patients with an elevated SCSA-defined sperm DNA fragmentation and prior ICSI failure(s).

Pregnancy Rates After Surgical Treatment of Deep Infiltrating Endometriosis in Infertile Patients With at Least 2 Previous In Vitro Fertilization or Intracytoplasmic Sperm Injection Failures

To assess the postoperative probabilities of pregnancy in patients with deep infiltrating endometriosis (DIE) and ≥2 previous in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) failures. Retrospective study using data prospectively recorded in the North-West Inter Regional Female Cohort for Patients with Endometriosis (CIRENDO) database. University tertiary referral center. Infertile patients under the age of 43 years, having undergone ≥2 previous IVF or ICSI failures, who were surgically managed for DIE. Complete excision of DIE. The pregnancy rate after surgery was assessed. One hundred and four infertile patients had surgery in 7 different centers participating in the database. Seventy-seven women intended to get pregnant postoperatively. Four patients who got pregnant by oocyte donation were excluded, resulting in a sample of 73 women. The mean patient age was 31.9 years (standard deviation [SD], 4.1), and the mean length of history of infertility was 48.4 months (SD, 26.5). Stage III and IV endometriosis were recorded in 83.6% of patients. The mean postoperative follow-up was 46.6 months (SD, 20.5). The postoperative pregnancy rate was 43.8% with a mean time from surgery to pregnancy of 11.1 months. 21.8% of pregnancies were spontaneous, 31.2% were obtained by IVF, 21.8% by frozen embryo transfer, 18.7% by IVF-ICSI, and 3.1% by intrauterine insemination. Multivariate analysis revealed that ovarian surgery, age ≥35 years old, and stage II endometriosis was associated with the probability of conception. Infertile women with ≥2 IVF-ICSI failures may be referred for surgery as it appears related to reasonable postoperative pregnancy rates, particularly when endometriomas surgery is either not required or not performed. Surgery for DIE does not routinely delay conception, as it usually occurs during the year following surgery.

Sperm chromatin condensation defects, but neither DNA fragmentation nor aneuploidy, are an independent predictor of clinical pregnancy after intracytoplasmic sperm injection.

The impact of sperm DNA damage on intracytoplasmic sperm injection (ICSI) outcomes remains controversial. The purpose of the study was to evaluate the prognostic value of several types of sperm nuclear damage on ICSI clinical pregnancy. Our retrospective study included a total of 132 couples who consulted for male or mixed-factor infertility that benefited from ICSI cycles from January 2006 to December 2015. All infertile males presented at least one conventional semen parameter alteration. Sperm nuclear damage was assessed using the Motile Sperm Organelle Morphological Examination for sperm head relative vacuolar area (RVA), aniline blue staining for chromatin condensation, terminal deoxynucleotidyl transferase dUTP nick-end labeling for DNA fragmentation, and fluorescence in situ hybridization for aneuploidy. Infertile males who achieved pregnancy after ICSI had fewer chromatin condensation defects than did males who did not achieve any pregnancy (15.8 ± 12.0% vs. 11.4 ± 7.9%, respectively, P = 0.0242), which remained significant in multivariate regression analysis (RR = 0.40 [0.18 to 0.86], P = 0.02). RVA, DNA fragmentation, and aneuploidy were not predictive factors of ICSI outcomes. The pregnancy rate was significantly decreased by number of progressive motile spermatozoa with normal morphology after migration (P = 0.04). In female partners, 17β estradiol of less than 2000pg/mL on the day of ovulation induction significantly reduced the occurrence of clinical pregnancy (P = 0.04). Sperm chromatin condensation defects were more frequently observed in couples with ICSI failure and should be considered a negative predictive factor for the occurrence of clinical pregnancy.

Assessment of CD56+ CD69+ Natural Killer Cells in the Peripheral Blood of Women with Repeated Implantation Failure (RIF) undergoing an Intra Cytoplasmic Sperm Injection (ICSI) Cycle

Background: infertility is an increasing medical and social problem. Delayed childbearing exposes couples to increased risk of causes of infertility including endometriosis, pelvic infection, and poor egg and sperm quality. Many infertile couples turn to in-vitro fertilization (IVF). Objective: to analyze and compare between the peripheral blood NK cells (CD56+ CD69+ ) in women with repeated ICSI failure. Patients and Methods: the current study is a case control study. One hundred women were included and divided into two groups: The study group included 50 women with history of unexplained infertility and repeated implantation failure in their ICSI attempts and a control group which included 50 normal, non-pregnant and fertile women. Results: the current study revealed that there was a highly statistically significant difference between control and case groups regarding CD3- CD56+dim NK percentage with a p-value <0.0001. The current study revealed that there was a statistically significant difference between control and case groups regarding CD3- CD56+dim CD69+ NK cells percentages with a p-value 0.001. Conclusion: there was a highly statistically significant difference between control and case groups regarding CD3- CD56+dim NK percentage. Also the study showed that total CD56+dim NK had a very good predictive value as it had a sensitivity of 80.0%, a specificity of 66.7%, a PPV of 73.3% and a NPV of 78.8%.

Artificial oocyte activation improves cycles with prospects of ICSI fertilization failure: a sibling oocyte control study.

Does artificial oocyte activation improve clinical outcomes for patients at risk of intracytoplasmic sperm injection (ICSI) fertilization failure? In this study, sibling oocytes from patients with previous ICSI failure or severe teratozoospermia were divided equally into two groups, half for artificial oocyte activation (AOA) with ionomycin after conventional ICSI and the other half for conventional ICSI only (non-AOA). The fertilization rates, cleavage rates, transferable embryo rates and blastulation rates of the two groups were compared first; the clinical pregnancy and live birth rates were also compared to assess the efficiency and safety of AOA. The outcomes of the AOA group were significantly better than those of the conventional ICSI group in terms of the fertilization (50.38% versus 33.86%, respectively, P<0.001), cleavage (59.16% versus 39.04%, respectively, P<0.001) and transferable embryo rates (43.51% versus 26.69%, respectively, P<0.001). The blastulation (43.53% versus 36.11%, respectively), implantation (26.83% versus 15.79%, respectively), clinical pregnancy (38.46% versus 25%, respectively) and live birth rates (38.46% versus 16.67%, respectively) were not significantly different. This study showed that AOA improved some aspects of cycles at risk of ICSI failure by increasing the fertilization and transferable embryo rates. But blastulation, pregnancy and implantation rates were not improved. The study is limited by its small size and absence of data on cumulative outcomes.

DNA fragmentation index and human papilloma virus in males with previous assisted reproductive technology failures.

This study was designed to evaluate the prevalence of Human Papilloma Virus (HPV) in semen and document the cycle outcomes in couples with previous intra-cytoplasmic sperm injection (ICSI) failures. One hundred and seventeen couples with at least two ICSI attempts were included in the study. HPV infection in semen and DNA fragmentation in samples were analyzed by commercially available kits. The percentage of spermatozoa with fragmented DNA (DNA fragmentation index: DFI) was determined during fluorescence microscopic examination as previously described. The cycle outcomes of couples with or without HPV infected male partners were recorded. According to our results, the prevalence of HPV was 7.7% in asymptomatic males with at least two previous ICSI failures. The increased DFI (>30%) was observed in 82.9% of the cases. In HPV-positive cases significantly lower number of good quality embryos were obtained. The implantation and pregnancy rates were similar in infected and non-infected males (p>0.05). The early miscarriage rate was slightly higher in HPV- positive group (33% vs. 10%, p>0.05). In cases with previous ICSI failures, the prevalence of HPV infection in semen is not higher than previously reported infertile populations. The reproductive outcome might be impaired in HPV-positive semen due to lower number of good quality embryos, which needs to be clarified by further large population-based studies.

Effect of endometrial scratch using a Pipelle catheter on recurrent implantation failure with intracytoplasmic sperm injection

Back ground In the last two decades, assisted reproductive technology procedures for culturing and transferring embryos have been continually improved, but the clinical pregnancy rate has not improved substantially over the last 10 years. The most important cause for implantation failure is embryo quality or uterine receptivity, but remains unexplained in many cases. About 75% of conceptus are lost before or at the time of implantation. Aim This work aims to assess the effect of endometrial scratch (ES) using a Pipelle catheter on the outcome of intracytoplasmic sperm injection (ICSI) in patients with recurrent implantation failure. Patients and methods The current study included 130 patients who were divided into two equal groups of 65 patients each. In the scratch group, endometrial local scratch using a Pipelle biopsy catheter on the mid luteal phase of spontaneous menstrual cycle before the ICSI procedure was performed, whereas the control group did not undergo ES. ICSI failure was recorded two or more times in each group. Results In the current study, there were no statistically significant differences between the two groups in age, gravidity and parity, type, duration of infertility, and the number of previous ICSI trials. However, the BMI was higher in the control group. In the current study, there were no statistically significant differences between the two groups in basal hormonal profile follicle-stimulating hormone, luteinizing hormone, thyroid-stimulating hormone, prolactin, estradiol, and anti-Mullerian hormone. In this study, there was highly significant increase in the pregnancy rate in the scratch group than the nonscratch group. The number of pregnant cases in the scratch group was 36 (55.4%), whereas in the control group, it was 21 (32.3%), with a highly significant P value of 0.008. In this study, there was no statistically significant difference between positive and negative pregnancy in the basal hormonal profile, except anti-Mullerian hormone and follicle-stimulating hormone levels, P values of 0.040 and 0.000. Conclusion ES in the cycle before the ICSI cycle improved the chemical and clinical pregnancy rates compared with ICSI performed without ES in cases of recurrent implantation failure. It seems that increasing pregnancy rates could be because of the local inflammatory reaction elicited by the endometrial sampling procedure.

A Familial Case Report of a 13;22 Chromosomal Translocation with Recurrent Intracytoplasmic Sperm Injection Failure.

The importance of cytogenetic analysis in a family with reproductive failure in two siblings is highlighted, where two siblings and their mother presented with a balanced translocation between chromosomes 13;22. The clinical evaluation had shown the female to be normal and the male to be oligoasthenoteratozoospermic despite repeated semen analysis. The couple was referred to our laboratory after three consecutive intracytoplasmic sperm injection (ICSI) failures at a local assisted reproductive technique (ART) center. Peripheral blood lymphocytes, obtained for karyotyping, were studied by a standard G-banding technique. Chromosomal analysis of the members of the pedigree, including the probands, showed the presence of the same translocation, t(13;22)(q21.2;q13.3), carried by three generations of the family. The sister and the mother of the proband had multiple spontaneous abortions in the first trimester. The spouses, when examined cytogenetically, were found to be normal. We propose the involvement of a balanced t(13;22)(q21.2;q13.3) chromosomal translocation in the pathogenesis of recurrent ART or spontaneous reproductive failures. Hence, it is suggested that all cases with structural chromosomal abnormalities be counseled prior to opting for ART and undergoing pre-implantation genetic diagnosis (PGD). This would prevent recurrent financial, physical and emotional stress in couples seeking ART.

Protocol shift from agonist to antagonist or vice versa after an unsuccessful intracytoplasmic sperm injection cycle on the same patient does not improve outcome

ObjectiveWhile there's a growing data on the comparison of GnRH agonist and GnRH antagonist protocols, no clear study exists on the effects of both protocols on the same patient. The aim of the present study was to compare the success rates of protocol shift and proceeding with the same protocol after an unsuccessful intracytoplasmic sperm injection (ICSI) cycle. Materials and MethodsThree hundred and forty-five normal responder patients who had a previously failed ICSI cycle between January 2011 and December 2015 were reviewed. The study (n = 73) and control (n = 93) groups in the first cohort were composed of patients whose protocol were shifted to antagonist and who proceeded with agonist. The study (n = 33) and control (n = 146) groups in the second cohort were composed of patients whose protocol were shifted to agonist and who proceeded with antagonist. ResultsTotal dose of gonadotropins, maximum estradiol levels, and number of oocytes collected were significantly higher in agonist protocol than in antagonist protocol (P = 0.021, P = 0.013, and P = 0.003, respectively). Cycle cancellation rates were significantly lower in agonist protocol than antagonist protocol (P = 0.005). The pregnancy rates in patients who shifted to antagonist and proceeded with agonist were 57.8% and 50.8%, respectively (P = 0.399). The pregnancy rates in patients who shifted to agonist and proceeded with antagonist were 33.3% and 47.9%, respectively (P = 0.202). ConclusionProtocol shift following a failed ICSI cycle with either agonist or antagonist protocol does not affect outcome.

Should A Couple with Failed In Vitro Fertilization/Intracytoplasmic Sperm Injection and Increased Sperm DNA Fragmentation Use Testicular Sperm for the Next Cycle?

Limited observational data purporting a benefit of testicular sperm for intracytoplasmic sperm injection failure do not provide sufficient evidence. The first step is to reduce sperm DNA fragmentation in ejaculated sperm through lifestyle modifications, antioxidant therapy, or treatment of underlying abnormalities such as infection or varicocele.

The Effect of Endometrial Injury on Pregnancy Rate in Women with Previous Intracytoplasmic Sperm Injection Failure/ a Randomized Clinical Trial

Local injury to the endometrium using a pipell endometrial sampler doubles the incidence of successful pregnanciesin patients undergoing in vitro fertilization. This study was confirmed by Local injury to endometriam of IVF patients in controlled ovarian hyperstimulation cycle may increase the incidence of embryo implantation.The successful pregnancy outcome of patients undergoing ovarian stimulation for IVF or related advanced reproductive technologies depend on several factors. Among these,embryo quality and intrauterine environment play a major role for achievement and further continuation of pregnancy.to review the effect of local injury to the endometrium on the outcome of ICSI and embryo transfer.This prospective observational study was conducted in ART unit at Ain Shams University Maternity Hospital during the period from July 2013 to July 2014. One hundered women,aged 23-40 years,with one or more failed ICSI cycle participated in this study. Nulligravidae represented 84% of all women in the study.The mean age was (30.38±4.41 years),the mean duration of marriage (7.54±4.95 years) and the mean duration of infertility (7.28±5.03 years),84% of patients were nulligravidae while 16% had history of abortions,none of our patients had previous full term pregnancy or live born children,88% of women involved in the study had a single failed trial while 12% had previous two failed trials. The highest percentage of cases showed normal hysteroscopic findings (52%),while endometritis showed the least percentage (4%),adhesions were found in (10%),polypi in (14%),congenital uterine malformation in (14%) and myoma in (4). 78% of cases had no intervention (only diagnostic hysteroscopy),while 22% of cases had operative interventions in the form of adhesiolysis (4%),polypectomy in (12%) and resection of a septum in (2%).The range of pain score was 2-10 with a mean (4.74±1.84min) while the range of duration of procedure was 1-25 with a mean (5.16±4.69 min).No statistically significant difference was found as regards the presence of abnormal hysteroscopic findings in relation to the number of failed trials (P&gt;0.05).There was no statistically significant difference between the two groups (diagnostic and operative groups) as regards the meanage,duration of marriage,duration of infertility and number of failed trials (P&gt;0.05). Operative hysteroscopy showed significantly longer duration than diagnostic (11.73 ±6.2 vs 3.26 ±1.29 min),respectively (P&lt;0.001).Cases of diagnostic hysteroscopy showed significantly lower VAS when compared to cases of operative hysteroscopy with a mean of (4.54 ± 1.42 vs 6.11 ± 2.6),respectively (P&lt;0.001),while operative hysteroscopy showed significantly higher VAS than diagnostic (P&lt;0.001).There was no statistically significant difference between hysteroscopic modality as regards the distribution of acceptability (P&gt;0.05),although acceptability of office hysteroscopy was higher in diagnostic group,yet the difference didn’t reach statistical significance.This technique has permitted complete elimination of any type of pre-medication,analgesia,or anesthesia,making the procedure faster,more acceptable and complication-free.In our study patient’s overall acceptability was 86%. The patient acceptability was non-significantly higher in diagnostic compared to operative hysteroscopy (89.7% vs 72.7%),respectively. As regards patients acceptability of office hysteroscopy,those who accepted the procedure showed significantly lower rate of pain and when compared to cases not accepting hysteroscopy (P&lt;0.05). There was a significant correlation between the pain and duration of the procedure (r = 0.35,P&lt;0.001). This signifies that the longer the duration of the procedure,the higher the pain in our study.Outpatient management is a comparatively new concept in gynecology. Patients may favor the outpatient approach because they spend less time in hospital. However,it is important to tailor this management according to patient status.

Can intracytoplasmic Morphologically Selected Spermatozoa Injection be Used as First Choice of Treatment for Severe Male Factor Infertility Patients?

Aim:This study was carried out to assess the outcome of the intracytoplasmic morphologically selected sperm injection (IMSI) technique compared with the previous failed intracytoplasmic sperm injection (ICSI) attempts in oligoasthenoteratozoospermia (OAT)/severe OAT (SOAT)/teratozoospermia patients.Setting:Institution-based, in vitro fertilization center.Study Design:It was a nonrandomized prospective study including 57 couples who had previous one or two ICSI failures (failure due to no implantation as embryos were transferred in these cycles and had no pregnancy) due to male factor. There was no case of total fertilization failure. IMSI was carried out in these couples and the results were compared with their previously failed ICSI attempts.Materials and Methods:Real-time selection of sperms was done using IMSI as it allows the assessment of fine nuclear morphology and vacuoles in the sperm head at a high magnification (>6000×) with differential interference contrast optics. Therefore, IMSI was applied in couples having OAT, SOAT or teratozoospermia as male factor and the results were compared with their previous failed ICSI attempts. Statistical analysis was carried out using GraphPad Prism.Results and Conclusion:Data analysis demonstrated significant difference in the fertilization rate between IMSI and previous ICSI attempts of these patients (30.0% vs. 52.0%; P < 0.05). The embryo quality, implantation and pregnancy rates with IMSI were also significantly higher than those of their previous ICSI cycles (32% vs. 56.4%; 30.2% vs. 68.5%; 0.0% vs. 62.4%; P < 0.05). Our conclusion is that the IMSI procedure improved embryo development and the clinical outcomes in the same infertile couples with male infertility and poor embryo development over their previous ICSI attempts and can be taken up as the treatment of choice in cases of severe male factor infertility.

Clinical analyses of successful and previously failed intracytoplasmic sperm injection cycle parameters in patients with poor ovarian reserve

Objective:To determine some major characteristic differences between two consecutive successful and unsuccessful intracytoplasmic sperm injection (ICSI) cycles in poor responders.Materials and Methods:Sixty women with poor ovarian response as determined using the Bologna criteria underwent ICSI cycles following an unsuccessful trial. Some parameters of both cycles including age, body mass index (BMI), serum follicle-stimulating hormone (FSH) and estradiol levels, antral follicle count, gonadotropin dosage, duration of stimulation, antagonist starting day, duration of antagonist administration, endometrial thickness at trigger day, number of total and fertilized oocytes, embryo transfer day, number of embryo cells, and fertilization rate were compared in the same patients to identify predictors of cycles with clinical pregnancy.Results:The mean age, BMI, serum FSH, estradiol concentrations, and antral follicle count were 35.9 years (range, 30-42 years), 25.9 kg/m2 (range, 18.4-33.5 kg/m2), 10.9 IU/mL (range, 7-13 IU/mL), 52.9 pg/mL (range, 11.6-75 pg/mL), and 4.7 (range, 2-10), respectively. A comparison of cycle characteristics showed a significantly higher total number of mature and fertilized oocytes in successful cycles. The fertilization rate was also significantly higher in cycles with clinical pregnancy. Early initiation of antagonist was shown to result in favorable outcomes. A comparison of embryo characteristics showed that transfer of higher-stage embryos and embryos with higher numbers of cells had a significant impact on cycle outcomes.Conclusion:Our comparison of parameters of failed and successful ICSI cycles in poor responders revealed significantly earlier antagonist initiation, higher total number of mature and fertilized oocytes, fertilization rate, and significantly higher stage of embryo development and cell numbers at transfer in cycles that resulted in clinical pregnancy.

SEPT12-NDC1 Complexes Are Required for Mammalian Spermiogenesis.

Male factor infertility accounts for approximately 50 percent of infertile couples. The male factor-related causes of intracytoplasmic sperm injection failure include the absence of sperm, immotile sperm, immature sperm, abnormally structured sperm, and sperm with nuclear damage. Our knockout and knock-in mice models demonstrated that SEPTIN12 (SEPT12) is vital for the formation of sperm morphological characteristics during spermiogenesis. In the clinical aspect, mutated SEPT12 in men results in oligozoospermia or teratozoospermia or both. Sperm with mutated SEPT12 revealed abnormal head and tail structures, decreased chromosomal condensation, and nuclear damage. Furthermore, several nuclear or nuclear membrane-related proteins have been identified as SEPT12 interactors through the yeast 2-hybrid system, including NDC1 transmembrane nucleoporin (NDC1). NDC1 is a major nuclear pore protein, and is critical for nuclear pore complex assembly and nuclear morphology maintenance in mammalian cells. Mutated NDC1 cause gametogenesis defects and skeletal malformations in mice, which were detected spontaneously in the A/J strain. In this study, we characterized the functional effects of SEPT12–NDC1 complexes during mammalian spermiogenesis. In mature human spermatozoa, SEPT12 and NDC1 are majorly colocalized in the centrosome regions; however, NDC1 is only slightly co-expressed with SEPT12 at the annulus of the sperm tail. In addition, SEPT12 interacts with NDC1 in the male germ cell line through coimmunoprecipitation. During murine spermiogenesis, we observed that NDC1 was located at the nuclear membrane of spermatids and at the necks of mature spermatozoa. In male germ cell lines, NDC1 overexpression restricted the localization of SEPT12 to the nucleus and repressed the filament formation of SEPT12. In mice sperm with mutated SEPT12, NDC1 dispersed around the manchette region of the sperm head and annulus, compared with concentrating at the sperm neck of wild-type sperm. These results indicate that SEPT12–NDC1 complexes are involved in mammalian spermiogenesis.

Effects of sperm quality on the success of intracytoplasmic sperm injection (ICSI) with testicular sperm in couples with recurrent ICSI failure with ejaculated sperm

Effects of sperm quality on the success of intracytoplasmic sperm injection (ICSI) with testicular sperm in couples with recurrent ICSI failure with ejaculated sperm

Rescue of Failed Oocyte Activation After ICSI in a Mouse Model of Male Factor Infertility by Recombinant Phospholipase Cζ

Artificial oocyte activation to overcome failed fertilization after intracytoplasmic sperm injection (ICSI) in human oocytes typically employs Ca2+ ionophores to produce a single cytosolic Ca2+ increase. In contrast, recombinant phospholipase Czeta (PLCζ) causes Ca2+ oscillations indistinguishable from those occurring during fertilization, but remains untested for its efficacy in a scenario of ICSI fertilization failure. Here, we compare PLCζ with other activation stimuli in a mouse model of failed oocyte activation after ICSI, in which heat-treated sperm are injected into mouse oocytes. We show that increasing periods of 56°C exposure of sperm produces a progressive loss of Ca2+ oscillations after ICSI. The decrease in Ca2+ oscillations produces a reduction in oocyte activation and embryo development to the blastocyst stage. We treated such oocytes that failed to activate after ICSI either with Ca2+ ionophore, or with Sr2+ media which causes Ca2+ oscillations, or we injected them with recombinant human PLCζ. All these treatments rescued oocyte activation, although Sr2+ and PLCζ gave the highest rates of development to blastocyst. When recombinant PLCζ was given to oocytes previously injected with control sperm, they developed normally to the blastocyst stage at rates similar to that after control ICSI. The data suggest that recombinant human PLCζ protein is an efficient means of rescuing oocyte activation after ICSI failure and that it can be effectively used even if the sperm already contains endogenous Ca2+ releasing activity.

Rescue of failed oocyte activation after ICSI in a mouse model of male factor infertility by recombinant phospholipase Cζ.

Artificial oocyte activation to overcome failed fertilization after intracytoplasmic sperm injection (ICSI) in human oocytes typically employs Ca2+ ionophores to produce a single cytosolic Ca2+ increase. In contrast, recombinant phospholipase Czeta (PLCζ) causes Ca2+ oscillations indistinguishable from those occurring during fertilization, but remains untested for its efficacy in a scenario of ICSI fertilization failure. Here, we compare PLCζ with other activation stimuli in a mouse model of failed oocyte activation after ICSI, in which heat-treated sperm are injected into mouse oocytes. We show that increasing periods of 56°C exposure of sperm produces a progressive loss of Ca2+ oscillations after ICSI. The decrease in Ca2+ oscillations produces a reduction in oocyte activation and embryo development to the blastocyst stage. We treated such oocytes that failed to activate after ICSI either with Ca2+ ionophore, or with Sr2+ media which causes Ca2+ oscillations, or we injected them with recombinant human PLCζ. All these treatments rescued oocyte activation, although Sr2+ and PLCζ gave the highest rates of development to blastocyst. When recombinant PLCζ was given to oocytes previously injected with control sperm, they developed normally to the blastocyst stage at rates similar to that after control ICSI. The data suggest that recombinant human PLCζ protein is an efficient means of rescuing oocyte activation after ICSI failure and that it can be effectively used even if the sperm already contains endogenous Ca2+ releasing activity.

SEPT12/SPAG4/LAMINB1 Complexes are Required for Maintaining the Integrity of the Nuclear Envelope in Postmeiotic Male Germ Cells

Male infertility affects approximately 50% of all infertile couples. The male-related causes of intracytoplasmic sperm injection failure include the absence of sperm, immotile or immature sperm, and sperm with structural defects such as those caused by premature chromosomal condensation and DNA damage. Our previous studies based on a knockout mice model indicated that SEPT12 proteins are critical for the terminal morphological formation of sperm.

Relationship between sperm parameters and intracytoplasmic sperm injection outcome

Abstract Objectives: With the adventure of intracytoplasmic sperm injection (ICSI) technique, great progresses have developed in the treatment of infertility. Concentration on the properties of male’s gamete has been encouraged by the increasing concerns about the causes of ICSI failure. We hence conducted this study to investigate the probable association of sperm parameters with ISCI outcome. Methods: A total of 523 couples referred to Isfahan Fertility and Sterility Center from January 2007 to June 2008 for ICSI. Semen analysis was performed before ICSI procedure according to the WHO criteria. Patients were assigned into successful ICSI (case) and failed ICSI (control) groups. Sperm parameters were then compared between the 2 groups. Results: One hundred and six patients (20%) had successful ICSI results (case group) compared with 417 couples (80%) with undesirable ICSI outcomes (control group). Among evaluated factors, sperm agglutination ( p = 0.007), sperm concentration ( p = 0.043), leukocytospermia ( p = 0.026) and head abnormality of sperm ( p = 0.019) showed statistically significant differences between two groups with differing ICSI results. None of the other semen parameters revealed significant differences between these two groups. Conclusion: Our study showed that some sperm parameters are associated with desirable ICSI outcome. However, it is unclear whether these associations are causal.

Evaluation of sperm nuclear integrity in patients with different percentages of decapitated sperm in ejaculates

The decapitated sperm defect is a rare type of teratozoospermia responsible for male infertility. Spermatozoa from patients affected by this syndrome are used for intracytoplasmic sperm injection (ICSI) although little is known about their DNA integrity. This study evaluated sperm nuclear alterations in four patients and ten fertile men (control group). Sperm samples were examined by light, transmission electron and high-magnification contrast microscopy and analysed after terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling, aniline blue staining and fluorescence in-situ hybridization. Spermatozoa from patients presented varying degrees of decapitation, along with morphological and ultrastructural head abnormalities. Whereas the proportion of spermatozoa with fragmented DNA and numerical chromosome abnormalities was similar in patients 1–3 and controls, the percentage of spermatozoa with hypocondensed chromatin was higher in patients 1–3 than in fertile men. Patient 4 presented a distinct phenotype, with an increased proportion of flagellated spermatozoa with DNA strand breaks as well as increased aneuploidy and diploidy rates compared with controls and with patients 1–3. No successful pregnancy resulted from ICSI although embryos were obtained for three patients. The morphological defects and the nuclear alterations observed in spermatozoa of patients with the decapitated sperm syndrome may have contributed to ICSI failures.