Effect Of Intracytoplasmic Sperm Injection Research Articles

STANDARD INSEMINATION (SI) IN IVF PATIENTS WITHOUT MALE FACTOR INFERTILITY (MFI) MAY INCREASE THE EUPLOIDY RATE (ER) AMONG PREIMPLANTATION GENETICALLY TESTED BLASTOCYSTS FOR ANEUPLOIDIES (PGT-A)

Available publications on the effects of intracytoplasmic sperm injection (ICSI) on ERs demonstrate conflicting outcomes.1,2 We aim to explore a large academic practices’ use of ICSI among patients with normospermia undergoing a controlled ovarian hyperstimulation in vitro fertilization (COH IVF) cycle with a trophectoderm biopsy for PGT-A.

The effect of oocyte denudation time and intracytoplasmic sperm injection time on embryo quality at assisted reproductive technology clinic – A cross-sectional study

BackgroundThis study aims to determine whether there is an effect of oocyte denudation time and ICSI time on embryo quality at assisted reproductive Technology clinic. MethodsAn observational analytic retrospective study was conduct using cross-sectional study. The subject were oocytes from in-vitro fertilization procedures using the ICSI method at the assisted reproductive technology clinic in a private hospital in Bandung for the period 2017 - 2019. Three variables were oocyte denudation time, ICSI time and embryo quality collected from samples that met the research criteria. Data will be grouped based on denudation time (T1) and Intracytoplasmic Sperm Injection time (T2). ResultFrom the 5 groups of denudation time; 3–4 hours, 4–5 hours, 5–6 hours, 6–7 hours and more than 7 hours group, the denudation time of 3–4 hours group showed the highest number (66.7%) for excellent embryo quality results, while denudation time of more than 7 hours showed the lowest number (29.2%) for excellent embryo quality results with p < 0.001. From these figures, it shows that the ICSI time of 3–4 hours is superior to the ICSI time of 2–3 hours because the outcome of excellent embryo quality is higher while the outcome of poor embryo quality is lower, although the difference is not significant (p = 0.140). ConclusionThis study shows there was a significant difference in the effect of oocyte denudation time on embryo quality at assisted reproductive technology clinic. There was no significant difference in the effect of intracytoplasmic sperm injection (ICSI) time on embryo quality at assisted reproductive technology clinic.

Intracytoplasmic sperm injection vs. conventional in vitro fertilization in patients with non-male factor infertility

To compare the cumulative live birth rates (CLBRs) and cost effectiveness of intracytoplasmic sperm injection (ICSI) and conventional invitro fertilization (cIVF) for non-male factor infertility. A retrospective cohort study. Society for Assisted Reproductive Technology clinics. A total of 46,967 patients with non-male factor infertility with the first autologous oocyte retrieval cycle between January 2014 and December2015. None. The primary outcomes were CLBR, defined as up to 1 live birth from an autologous retrieval cycle between 2014 and 2015, and linked fresh and frozen embryo transfers through 2016. The secondary outcomes included miscarriage rate, 2 pronuclei per oocyte retrieved, and the total number of transferred and frozen embryos. Analyses were performed on subsamples with and without preimplantation genetic testing for aneuploidy (PGT-A). A cost analysis was performed to determine the costs accrued by ICSI. Among cycles without PGT-A in patients with non-male factor infertility, the CLBR was 60.9% for ICSI cycles vs. 64.3% for cIVF cycles, a difference that was not significantly different after adjustment for covariates (adjusted risk ratio, 0.99; 95% confidence interval, 0.99-1.00). With PGT-A, no difference in CLBR was found between ICSI and cIVF cases after adjustment (64.7% vs. 69.0%, respectively; adjusted risk ratio, 0.97; 95% confidence interval, 0.93-1.01). The patients were charged an estimated additional amount of $37,476,000 for ICSI without genetic testing and an additional amount of $7,213,500 for ICSI with PGT-A over 2 years by Society for Assisted Reproductive Technology clinics. In patients with non-male factor infertility, ICSI did not improve CLBR. Given the additional cost and the lack of CLBR benefit, our data show that the routine use of ICSI in patients with non-male factor infertility is not warranted.

Intracytoplasmic Sperm Injection Improves Normal Fertilization Rate and Clinical Pregnancy Rate in Male Infertility.

The purpose of this study was to investigate the effect of intracytoplasmic sperm injection (ICSI) on the normal fertilization rate of oocytes and the clinical pregnancy rate of cycles in male infertility. Eighty cases of male infertility patients attending our hospital from March 2020 to March 2021 were selected and divided into observation group and control group using the random number table method, with 40 cases in each group. The control group was treated with in vitro fertilization (IVF), and the observation group was treated with ICSI. The normal fertilization rate of oocytes and the clinical pregnancy rate of the cycle were compared between the two groups, and the adverse pregnancy outcome and obstetric and perinatal complications were compared between the two groups The rate of normal fertilization and clinical pregnancy in the cycle was higher in the observation group (P < 0.05). The differences were not statistically significant (P > 0.05) when comparing adverse pregnancy outcomes between the two groups. The difference was not statistically significant (P > 0.05) when comparing obstetrics (5.41%, 10.34%) and perinatal complications (8.33%, 14.81%) between the two groups. ICSI in male infertility is significantly effective in improving the rate of normal oocyte fertilization and the clinical pregnancy rate of the cycle. It also has a low impact on adverse pregnancy outcomes and obstetric and perinatal complications and has a high safety profile.

Does intracytoplasmic sperm injection increase the risk of gestational diabetes in patients with polycystic over?

Aim: Some studies found that in vitro fertilization (IVF) pregnancies were correlated with an increase in the incidence of gestational diabetes mellitus (GDM). The purpose of this study was to determine the effect of Intracytoplasmic Sperm Injection (ICSI) on the risk of GDM in patients with polycystic over syndrome (PCOS). Material and Methods: This retrospective study was conducted on 862 women below 40 who applied to gynecology and obstetrics clinic between January 2015 and May 2020. Their BMI was close to each other. They all had a single fetus, and they did not have a diabetes history. The individuals who participated in the study were tested based on 75 g OGTT (oral glucose tolerance test). Demographical characteristics, biochemical findings, and treatment styles were evaluated. Those with GDM history in the previous pregnancy were excluded from the study. Results: The mean age and BMI of the patients and the differences observed in the control and case groups were not statistically significant (p&amp;gt;0,05). Preparandial (p=0,000), 1st-hour blood glucose levels of OGTT (p=0,000), and 2nd-hour blood glucose levels of OGTT (p=0,000) of the case group were statistically higher than those in the control group (p&amp;lt;0,05). Conclusion: The confirmation of the indicators which increase the risk of GDM development with PCOS and follow-up in the early pregnancy period might minimize maternal and fetal complications depending on GDM. This study is significant since it investigates the effect of ICSI in patients with PCOS, spontaneous, and IVF pregnancies.

The effect of ICSI in infertility couples with non-male factor: a systematic review and meta-analysis.

We performed a systematic review and meta-analysis of available literature to investigate the efficacy of the intracytoplasmic sperm injection (ICSI) in couples with non-male factor with respect to the clinical outcomes. The literature search was based on EMBASE, PubMed, and the Cochrane Library. All studies published after 1992 until February 2020 and written in English addressing patients in the presence of normal semen parameters subjected to ICSI and in vitro fertilization (IVF) were eligible. Reference lists of retrieved articles were hand-searched for additional studies. The primary outcomes were fertilization rate, clinical pregnancy rate, and implantation rate; the secondary outcomes were good-quality embryo rate, miscarriage rate, and live birth rate. Four RCTs and twenty-two cohort studies fulfilling the inclusion criteria were included. Collectively, a meta-analysis of the outcomes in RCTs showed that compared to IVF, ICSI has no obvious advantage in fertilization rate (RR = 1.16, 95% CI: 0.83-1.62), clinical pregnancy rate (RR = 1.04, 95% CI: 0.66-1.64), implantation rate (RR = 1.12, 95% CI: 0.67-1.86), and live birth rate (RR = 1.17, 95% CI: 0.43-3.15). Pooled results of cohort studies demonstrated a statistically significant higher fertilization rate (RR = 1.16, 95% CI: 1.03-1.31) and miscarriage rate (RR = 1.04, 95% CI: 1.01-1.06) in the ICSI group; furthermore, higher clinical pregnancy rate (RR = 0.85, 95% CI: 0.77-0.94), implantation rate (RR = 0.78, 95% CI: 0.65-0.95), and live birth rate (RR = 0.86, 95% CI: 0.79-0.94) was founded in the IVF group; no statistically significant difference was observed in good-quality embryo rate (RR = 0.98, 95% CI: 0.93-1.04). ICSI has no obvious advantage in patients with normal semen parameters. Enough information is still not available to prove the efficacy of ICSI in couples with non-male factor infertility comparing to IVF.

Effects of testicular sperm aspiration upon first cycle ICSI-ET for type 2 diabetic male patients

ABSTRACT Male diabetes mellitus (DM) can affect erectile function and sperm quality. In severe cases, DM can lead to retrograde or no ejaculation, so testicular sperm aspiration (TESA) is combined with intracytoplasmic sperm injection (ICSI) to treat subfertility and infertility for DM couples. However, the effect of TESA upon ICSI (TESA-ICSI) for DM patients remains unclear. This research investigated the effect of TESA-ICSI on first cycle ICSI-embryo transfer (ICSI-ET) for type 2 diabetic mellitus (T2DM) patients and the potential mechanisms. The subjects consisted of 1219 male patients with azoospermia or retrograde ejaculation who were treated with TESA-ICSI from 2015.01 to 2019.11. They were classified into two groups, the T2DM group (n = 54) and non-diabetic control group (n = 1165). Sperm selection for injection was performed using motile sperm organelle morphology examination criteria. The number of available embryos and the high-quality embryo rates following a single ET as well as cleavage, fertilization, implantation, clinical pregnancy and the abortion rates were noted. Compared with the non-diabetic group, the available embryo rate (75.20 ± 26.40% vs.78.36 ± 23.25%) and high-quality embryo rate (46.49 ± 30.37% vs. 47.55 ± 28.57%) in the T2DM group were lower and the abortion rate (20.83% vs. 8.88%) was higher, but these differences were not statistically significant. There were no significant differences in clinical pregnancy, implantation, normal fertilization, and cleavage rates between the two groups. The results show that TESA for male T2DM patients does not influence the effect of ICSI. For T2DM patients with severe oligozoospermia, asthenospermia, teratozoospermia, or retrograde ejaculation that do not meet ICSI criteria, TESA-ICSI may perhaps be considered for reproductive assistance. Abbreviations DM: diabetes mellitus; TESA: testicular sperm aspiration; ICSI: intracytoplasmic sperm injection; ICSI-ET; ICSI-embryo transfer; LH: luteinizing hormone; mL: milliliter; TES: testosterone; FSH: follicle-stimulating hormone; P: progesterone; HCG: human chorionic gonadotropin

Comparing fertilization rates from intracytoplasmic sperm injection to conventional in vitro fertilization among women of advanced age with non−male factor infertility: a meta-analysis

To evaluate the effectiveness of intracytoplasmic sperm injection (ICSI) in improving fertilization rates compared to conventional invitro fertilization rates (IVF) among women aged ≥38 years with a non-male factor diagnosis. Systematic review and meta-analysis. Not applicable. Women aged ≥38 years with a non-male factor diagnosis receiving IVF or ICSI. A systematic review of databases including PubMed and Embase was performed. Study protocol was registered at the International Prospective Register of Systematic Reviews. Studies were selected if they compared fertilization rates from ICSI with those from conventional IVF among women aged ≥38 years with a non-male factor infertility diagnosis. A random effects model was used. Meta-analysis of Observational Studies in Epidemiology guidelines were applied. Fertilization rate. Seven studies including 8796 retrieved oocytes (ICSI: 4,369; IVF: 4,427) with mean female age ≥38 years met the inclusion criteria. There was no significant difference in fertilization rates between ICSI and conventional IVF (relative risk [RR] 0.99, 95% confidence interval [CI] 0.93-1.06; P = .8). Heterogeneity was observed between studies (I2 = 58.2; P < .05). Heterogeneity was significant (I2 = 57.1; P < .05) when cycles with prior fertilization failure were excluded; however, when analysis was restricted to poor responders (RR 1.01, 95% CI 0.97-1.05; P = .6), heterogeneity was no longer significant (I2 = 0.0; P = .5). No difference was found in fertilization rates between conventional IVF and ICSI. Further studies are needed to assess the impact of ICSI in this population, controlling for other indications such as preimplantation genetic testing.

Failure of complete hatching of ICSI-derived human blastocyst by cell herniation via small slit and insufficient expansion despite ongoing cell proliferation.

To assess the effect of intracytoplasmic sperm injection (ICSI) on embryo hatching and visualise the effects of zona thinning (ZT) on the embryo using time-lapse monitoring. In vitro fertilisation (IVF) (n = 178) and ICSI (n = 110)-derived cryopreserved blastocysts were donated by patients who previously had a baby. This study investigated the impacts of IVF, ICSI, laser-assisted hatching by ZT and formation of ICSI penetration trace on zona pellucida of IVF-derived blastocyst on blastcyst diameter, the estimated number of trophectoderm (TE) cells and completed hatching rate. The completed hatching rate and diameters of the completely hatched blastocysts at hatching commencement and at the maximum expansion were significantly greater in the IVF than in ICSI groups. The completed hatching rate significantly increased with ZT in both groups. The maximum diameters of the completely hatched blastocysts were significantly smaller in the ZT than in non-ZT groups. The estimated TE cell numbers increased from hatching commencement to their maximum expansion points. The incompletely hatched ICSI-derived blastocysts intermittently herniated cells via small slits until degeneration. The completed hatching rate significantly decreased by the formation of ICSI penetration trace on zona pellucida of IVF-derived blastocyst. ICSI-derived blastocysts intermittently release proliferating cells and extracted TE cells and/or inner cell masses via a small slit; thus, blastocyst expansion is not sufficiently increased, leading to a reduced complete hatching rate. Therefore, the ICSI penetration trace potentially has negative effects on blastocyst expansion process in vitro and is a risk factor for the failure of completed hatching.

142 POSSIBLE BENEFIT OF INTRACYTOPLASMIC SPERM INJECTION ON THE EFFICIENCY OF EMBRYO PRODUCTION IN AGED COWS

Reduction in oocyte quality is a major factor responsible for declining fertility with age. The abnormal fertilization rate of oocytes from aged cows was reportedly higher than that of oocytes from young cows (Iwata et al. 2011. Reprod. Fertil. Dev. 23, 424–432). We hypothesised that assisted fertilization by intracytoplasmic sperm injection (ICSI) might improve the developmental abilities of oocytes collected from aged females. The aims of the study were (1) to determine the effect of maternal aging on the ability of bovine oocytes to undergo cytoplasmic maturation, fertilization, and further embryo development; and (2) to determine whether ICSI would improve the efficiency of embryo production in aged cows. Cows aged 30 to 50 months or &gt;120 months were defined as young or aged, respectively. Cumulus-oocyte complexes were harvested from abattoir-derived ovaries of young (40 ± 7 months, n = 89) and aged (136 ± 12 months, n = 55) Holstein cows and matured for 23 h in TCM-199 supplemented with 5% fetal bovine serum (FBS) at 38.5°C under 5% CO2 with saturated humidity. Then, surrounding cumulus cells were removed, and cortical granules in the oocyte were stained with Lens culinaris–fluorescein isothiocyanate to evaluate the cytoplasmic maturation. Matured oocytes were inseminated by IVF or ICSI. At 15 h post-insemination, the numbers of pronuclei were determined to evaluate the fertilization rates. Presumptive IVF- or ICSI-derived zygotes were cultured for 5 days in CR1aa medium with 2% FBS and subsequently in USU6 with 5% FBS for 3 days at 38.5°C in 5% O2, 5% CO2, and 90% N2 with saturated humidity. Chromosome numbers of blastocysts were counted to evaluate the effect of maternal aging on ploidy. All experiments were performed with more than 4 independent runs, and data were analysed using chi-square tests. The distribution of matured oocytes into different cortical granule classes was affected by age, with a significantly lower (P &lt; 0.01) proportion of class III (mature cytoplasm) oocytes from aged cows (29%) compared with those from young cows (57%). Although fertilization rates following IVF did not differ between the groups, the proportion of abnormal fertilization (more than 2 pronuclei) was 32% in the aged group: higher than in the young group (15%; P &lt; 0.01). The rates of cleaved embryos following IVF were the same among groups, but the rate of development to the blastocyst stage of oocytes from aged cows (38%) was significantly (P &lt; 0.05) lower than in those from young cows (52%). Moreover, the proportion of diploid blastocysts with 2 sets of chromosomes (2n = 60) was lower (47%) in the aged than in the young groups (75%; P &lt; 0.05). However, in the ICSI embryos, the rates of development to the blastocyst stage did not differ significantly between groups (young 36%; aged 43%). Thus, maternal aging might impair the cytoplasmic maturation of bovine oocytes, which could be associated with abnormal fertilization or low developmental competence. Our results also indicate possible beneficial effects of ICSI on the efficiency of embryo production in aged cows.

Comparison of reproductive outcome in oligozoospermic men with high sperm DNA fragmentation undergoing intracytoplasmic sperm injection with ejaculated and testicular sperm

To investigate the effectiveness of intracytoplasmic sperm injection (ICSI) using testicular sperm as a strategy to overcome infertility in men with high sperm DNA fragmentation (SDF). Prospective, observational, cohort study. Private IVF centers. A total of 147 couples undergoing IVF-ICSI and day 3 fresh ETs whose male partner has oligozoospermia and high SDF. Sperm injections were carried out with ejaculated sperm (EJA-ICSI) or testicular sperm (TESTI-ICSI) retrieved by either testicular sperm extraction (TESE) or testicular sperm aspiration (TESA). SDF levels were reassessed on the day of oocyte retrieval in both ejaculated and testicular specimens. Percentage of testicular and ejaculated spermatozoa containing fragmented DNA (%DFI) and clinical pregnancy, miscarriage, and live-birth rates. The %DFI in testicular sperm was 8.3%, compared with 40.7% in ejaculated sperm. For the TESTI-ICSI group versus the EJA-ICSI group, respectively, the clinical pregnancy rate was 51.9% and 40.2%, the miscarriage rate was 10.0% and 34.3%, and the live-birth rate was 46.7% and 26.4%. ICSI outcomes were significantly better in the group of men who had testicular sperm used for ICSI compared with those with ejaculated sperm. SDF was significantly lower in testicular specimens compared with ejaculated counterparts. Our results suggest that TESTI-ICSI is an effective option to overcome infertility when applied to selected men with oligozoospermia and high ejaculated SDF levels.

Embryo manipulation via assisted reproductive technology and epigenetic asymmetry in mammalian early development

The early stage of mammalian development from fertilization to implantation is a period when global and differential changes in the epigenetic landscape occur in paternally and maternally derived genomes, respectively. The sperm and egg DNA methylation profiles are very different from each other, and just after fertilization, only the paternally derived genome is subjected to genome-wide hydroxylation of 5-methylcytosine, resulting in an epigenetic asymmetry in parentally derived genomes. Although most of these differences are not present by the blastocyst stage, presumably due to passive demethylation, the maintenance of genomic imprinting memory and X chromosome inactivation in this stage are of critical importance for post-implantation development. Zygotic gene activation from paternally or maternally derived genomes also starts around the two-cell stage, presumably in a different manner in each of them. It is during this period that embryo manipulation, including assisted reproductive technology, is normally performed; so it is critically important to determine whether embryo manipulation procedures increase developmental risks by disturbing subsequent gene expression during the embryonic and/or neonatal development stages. In this review, we discuss the effects of various embryo manipulation procedures applied at the fertilization stage in relation to the epigenetic asymmetry in pre-implantation development. In particular, we focus on the effects of intracytoplasmic sperm injection that can result in long-lasting transcriptome disturbances, at least in mice.

125 INTRACYTOPLASMIC SPERM INJECTION (ICSI)-BASED MOUSE EMBRYO ASSAY: CHOICE OF EMBRYO CULTURE SYSTEM OUTWEIGHS THE EFFECT OF FERTILIZATION PROCEDURE ON EMBRYO DEVELOPMENT

Human embryo culture media, intended for assisted reproductive technologies (ARTs), are released for clinical use if they pass the mouse embryo assay (MEA). This assay prescribes that at least 70% of in vivo fertilized mouse 1-cell embryos form blastocysts, in order to grant the culture medium approval. In the fertility clinic, however, human embryos undergo more manipulation than their MEA counterparts through, for example, fertilization by intracytoplasmic sperm injection (ICSI); further, only a minority of the embryos transferred to the uterus goes on to establish gestations. In this context, we asked if the results of the MEA only depend on the type of in vitro culture, or are also affected by the method of fertilization. Superovulated B6C3F1 mouse oocytes were fertilized by ICSI using C57Bl/6 sperm. Pronuclear-stage eggs were allocated to four developmental environments: two ART culture protocols (HTF/MultiBlast, Irvine Scientific; ISM1/ISM2, Origio), standard mouse culture medium (KSOM(aa), made in-house) and the oviduct of pseudopregnant CD1 mice. As control for the invasive manipulation, pronuclear-stage eggs were generated by mating (B6C3F1 × C57Bl/6) and cultured in KSOM(aa) medium. Embryos were recovered from culture or from the CD1 uterus and scored for blastocyst formation at 96 h of development (Table 1). For these blastocysts, we determined the number of total, inner cell mass (ICM), and trophectoderm (TE) cells (Table 1) by confocal immunofluorescence microscopy (Schwarzer et al. 2012 doi:10.1093/humrep/des223). Our results show that ART culture protocols applied to mouse ICSI embryos are not equivalent in supporting blastocyst formation. Based on blastocyst rates, the ranking observed here after ICSI, reflects the ranking reported by us for IVF embryos (Schwarzer et al. 2012); that is, KSOM(aa) &gt; HTF/MultiBlast &gt; oviduct &gt; ISM1/2. This similarity suggests that the effect of in vitro culture on mouse development exceeds the effect of ICSI, provided gametes are of good quality. From the analysis of cell numbers, we note that while the ICM/TE ratios are not of easy interpretation, the absolute numbers of cells in the ICM draw a clear line between the environment of the oviduct and those of culture media. Irrespective of the ICM/TE ratio, only the oviduct environment secures 8 cells in the ICM (Table 1). Soriano and Jaenisch (1986 Cell 46, 19–29) reported that 8 cells of the ICM are set aside to give rise to the body of a mouse. In summary, the current MEA is a valuable assay to assess the quality of culture medium, however, its refinement is necessary to better model the adaptive properties of embryo culture when different methods of fertilization are applied. Until the MEA is extended into postimplantation development, as we advocate (Schwarzer et al. 2012), the absolute numbers of cells in the ICM may be a better gauge of embryo quality than the blastocyst rates. Table 1.Mouse embryo assay outcomes after ICSI

The effect of intracytoplasmic sperm injection (ICSI) on the timing of embryonic progression in cycles where oocytes were split into both ICSI and conventional insemination

To assess whether timing of blastulation was altered between oocytes that were fertilized with conventional insemination versus intracytoplasmic sperm injection (ICSI). To also evaluate the differences in fertilization rates (FR), overall blastulation rates (BR) and pregnancy rates (PR).

The effect of intracytoplasmic sperm injection on the early hCG level in twin pregnancies

Objectives: Since it has been suggested that intracytoplasmic sperm injection (ICSI) might impair the early development of embryos and twin pregnancies have physiologically different aspects from singleton pregnancies, we aimed to investigate the effect of ICSI on the human chrionic gonadotropin (hCG) levels in twin pregnancies. Methods: A total of 220 clinical pregnancies which underwent controlled ovarian hyperstimulation and in vitro fertilization/ICSI were analyzed retrospectively. Results: In twin pregnancies, the mean number of retrieved oocytes and estradiol (E2) levels on hCG day were significantly higher in ICSI group in spite of lower dose of gonadotropin. The hCG levels according to insemination method did not demonstrate any difference after adjustment for age of patient and partner, BMI, treatment regimens, serum E2 on hCG day, number of oocytes retrieved and date of sampling. Conclusions: The hCG values according to fertilization methods were not significantly different both in twin pregnancies with adjustment for confounders.

The effect of intracytoplasmic sperm injection on gender in IVF cycles in patients undergoing preimplantation genetic screening (PGS)

OBJECTIVE: In the past 2 decades, studies have suggested differences in sex ratio following IVF treatment. Recently, analysis showed a trend towards male embryos when transferring fresh blastocysts (JARG 2009). It has also been suggested that ICSI might be associated with the delivery of more females (F&S 2009). Our study's objective was to determine whether the use of ICSI in IVF cycles for PGS is associated with gender. We studied all the fertlized embryos that were biopsies in patient undergoing PGS.

376 FAILURE OF SPERM DECONDENSATION AFTER INTRACYTOPLASMIC SPERM INJECTION IN SWAMP BUFFALO (BUBALUS BUBALIS)

Intracytoplasmic sperm injection (ICSI) has been intensively used to examine the early events of gamete activation, but few studies have been reported for swamp buffalo. The first objective (Exp. 1) was to compare the developmental competence of oocytes after ICSI using either live or dead frozen-thawed spermatozoa. Matured oocytes were fertilized by ICSI using live (n = 148) or dead (n = 151) spermatozoa, followed by chemical activation using calcium ionophore (A23187) and cyclohexamide (CHX) in SOF medium. In vitro fertilization (n = 149) served as thecontrol. Cleavage rate was recorded on Day 2 and blastocyst formation rate was evaluated on Day 7. The second objective (Exp. 2) was to examine the effects of ICSI and activation regime on the decondensation of buffalo spermatozoa. A total of 148 matured oocytes were subjected to ICSI. The sperm-injected oocytes (n = 87) were then activated using the activation protocol as described in Exp. 1. The ICSI oocytes without activation (n = 61) and sham-injected oocytes with activation (n = 35) were used as controls. Nuclear changes of presumptive zygotes were mor- phologically evaluated for pronuclear formation using 4′,6-diamidino-2-phenylindole procedure and epifluorescent microscopy at 18 h post-ICSI. Statistical differences were determined among the groups using chi-square test. In Exp. 1, the results showed that the percentages of cleavage and blastocyst formation rate were 79.7, 77.0, and 41.6% and 33.8, 30.5, and 14.8% in the live sperm, dead sperm, and IVF groups, respectively. Embryo development rates did not significantly differ between ICSI groups; however, these rates were significantly higher than in the IVF group (P &lt; 0.05). In Exp. 2, the pronuclear formation rate was significantly higher in the ICSI with chemical (70.1%) and sham injection with chemical (60.6%) groups than in the ICSI without chemical group (3.2%; P &lt; 0.01). However, most of the presumptive zygotes with pronuclear formation from the ICSI with chemical activation group showed only intact sperm heads instead of the full male pronuclear formation. Our study suggests that the chemical activation directly affected the female pronuclear formation and embryo development but that it was not associated with the male pronuclear formation. It is postulated that ICSI oocytes that developed to cleavage and blastocyst stages underwent parthenogenesis after chemical activation. This work was supported by TRF-MAG (MRG-WII515S056) and CHE-TRF Senior Research Fund (RTA5080010).

A successful pregnancy and live birth after intracytoplasmic sperm injection with globozoospermic sperm and electrical oocyte activation

To present the effectiveness of intracytoplasmic sperm injection (ICSI) using globozoospermic sperm and assisted oocyte activation by electrical stimulation. A case report. A private IVF center in Japan. A man with globozoospermia. Acridine orange (AO) test, mouse oocyte activation test, and ICSI with electrical oocyte activation. Fertilization, pregnancy, and live birth. In the first ICSI attempt, neither of the two injected oocytes fertilized. Staining of the patient's sperm with AO showed that only 2.9% of the sperm emitted a green fluorescence at the characteristic round head (sperm with native DNA content). The mouse oocyte activation test using the roundheaded sperm showed that the normal fertilization rate was 78.9% when SrCl(2) was used for assisted oocyte activation; however, it was 6.0% without assisted oocyte activation. We confirmed that the sperm had defective ability to activate oocytes. In the second ICSI attempt, human oocytes were activated electrically with use of a single square direct current pulse after microinjection. All the seven injected oocytes fertilized normally, and two eight-cell embryos were transferred on day 3. Clinical pregnancy was confirmed, and a healthy girl weighing 2362 g was delivered at 37 weeks of gestation by cesarean section. This is the first successful outcome of ICSI using globozoospermic sperm and electrical oocyte activation. The electroactivation obviates the need for the use of potentially harmful drugs for activation.

The effectiveness of intracytoplasmic sperm injection combined with piezoelectric stimulation in infertile couples with total fertilization failure

To assess the effectiveness of intracytoplasmic sperm injection (ICSI) combined with piezoelectric stimulation in infertile couples with a history of total fertilization failure (TFF). Prospective controlled trial. Clinical IVF laboratory. Seventy-one couples undergoing ICSI on sibling oocytes having at least one previous ICSI attempt with TFF. ICSI or ICSI with piezoelectric activation. Fertilization rate. The patients were allocated to two groups: group I included 21 patients with only one previous TFF and group II included 50 patients with more than one previous TFF. Collectively, a total of 823 metaphase II (MII) oocytes were retrieved in 78 oocyte retrievals. In Group I, combined ICSI with piezoelectric stimulation was applied to 123/211 (58.2%) of MII oocytes (group IA), whereas standard ICSI procedure was applied to 88/211 (41.8%) of MII oocytes (group IB). The fertilization rate was 62% and 12% in group IA and group IB respectively. In group II, piezoelectric activation was applied in all 612 MII oocytes, of which 296 (48.3%) were fertilized. The rates for implantation and pregnancy/embryo transfer were obtained as 30.6% and 44.1%, respectively. Piezoelectric activation seems to improve IVF outcome in patients with previous TFF history.

Intracytoplasmic Sperm Injection for Male Factor Infertility: Effects on Gender of Offspring

Background: Concerns over the effect of intracytoplasmic sperm injection (ICSI) on offspring exist, partly because a genetic cause may be associated with male infertility. Theoretically, ICSI could affect the gender ratio of liveborn offspring of patients who undergo in vitro fertilization (IVF) with ICSI.