Intracytoplasmic sperm injection in cases with a history of in vitro fertilization failure

<abstract>Aim: To evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure. Methods: Two groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase Ⅱ oocytes: 0~30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound-guided oocyte collection was done 35~37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase Ⅱ oocytes were selected for microinjection. Results: Fertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 %±21.1 % vs. 73.9 %±19.2 %), cleavage rate (85.1 %±19.3 % vs. 82.7 %±22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups. Conclusion: ICSI is a valuable method for couples with a history of IVF failure. These patients may have a similar ICSI result as in severe male infertility.

A live birth of activated one-day-old unfertilized oocyte for a patient who experienced repeatedly near-total fertilization failure after intracytoplasmic sperm injection

Total or near-total fertilization failure after intracytoplasmic sperm injection （ICSl） is a rare event, but it occurs repeatedly because of sperm defects in activating oocyte. The case presents a successful pregnancy and live birth after calcium ionophore A23187 （A23187） activation on one-day-old unfertilized oocytes in a patient whose husband suffered oligoasthenoteratozoospermia, and who had experienced repeated near-total fertilization failure after ICSI. In the second ICSI cycle, only one oocyte was fertilized while nine were unfertilized. Oocyte activation with A23187 were performed on the one-day-old unfertilized oocytes after ICSI and resulted in fertilization and embryo transfer. A clinical pregnancy was achieved and a healthy baby was born. To our knowledge, this is the first reported case of a healthy birth after oocyte activation on the one-day-old unfertilized oocyte. This indicates that ＂rescue oocyte activation＂ on one-day-old unfertilized oocytes after ICSI may be helpful for preventing total or near-total fertilization failure after ICSI.

Related Factors of in vitro Fertilization and Embryo Transfer Patients with Complete Fertilization Failure

Objective To find the possible factors predicting fertilization failure of in vitro fertilization-embryo transfer （1VF-ET）. Methods The IVF-ET patients with complete fertilization failure （experimental group, n =32） were analyzed retrospectively. The patients whose oocytes retrieved at the same day and cultured on the same incubators with ≥ 50% fertilization rates were matched as the control （n=56）. Results The infertility duration, superovulation days, the rates of primary case, progesterone （P） level 〉3.12 nmol/L rate and rate of severe abnormal sperm （abnormal sperm rate 〉95%） in experimental group were significantly higher than those in the control （6.4 ±3.1 years, 12.6 ±2.2 d, 56%, 43%, 43% vs 4.6±2.9years, 11.6 ±% 1.3 d, 33%, 23%, 23%, respectively, P〈0.05）. Conclusion We should pay attention to these patients with primary infertility, longer infertility duration and superovulation days （〉6.4 years and 〉12.6 d） and having increased level of P on hCG injection day （〉3.12 nmol/L）, abnormal sperm rate 〉95% at the same time. They should be included in such patients at high risk of fertilization failure.

A homozygous protein-truncating mutation in ACTL7A causes male infertility characterized by fertilization failure

Objective:This study aimed to screen for novel mutations in ACTL7A and expand the spectrum of known mutations responsible for recurrent fertilization failure.Methods:Whole-exome sequencing was performed on samples from couples who experienced recurrent assisted reproductive technology failure and visited the General Hospital of Ningxia Medical University.Western blotting and quantitative Real-time PCR were used to investigate the effects of the mutation on HEK293T cells.Results:Samples from 12 couples with total fertilization failure or poor fertilization(fertilization rate<20%)were subjected to whole-exome sequencing,and a novel homozygous protein-truncating mutation(c.1101dupC,p.S368Qfs*5)in ACTL7A was identified in a patient with recurrent poor fertilization.The mutant resulted in a truncated protein as well as decreased protein expression level in HEK293T cells.Conclusions:Our findings expand the mutational and phenotypic spectrum of ACTL7A,thus providing a potential diagnostic marker for fertilization failure due to male factors.

Is There Any Role for Calcium Ionophore in ICSI Cycles for Cases of Non Obstructive Azoospermia?

Objectives: Evaluate the effect of artificial oocyte activation (AOA) using calcium ionophore (A23187) on the rate of fertilization and cleavage of embryos in surgically retrieved sperm of patients with non-obstructive azoospermia undergoing intracytoplasmic sperm injection (ICSI). Study design: This study was conducted on 60 infertile couples undergoing ICSI cycles as a randomized controlled parallel group’s experimental study in a private IVF center in Egypt from January 2018 to July 2019. ICSI cycles were divided into two groups: Group A: includes 30 ICSI patients with surgically retrieved sperms of non-obstructive azoospermia treated with calcium ionophore (A23187). Group C/Control: includes 30 ICSI patients with surgically retrieved sperms of non-obstructive azoospermia non-treated with calcium ionophore (A23187). Results: There was no statistical difference between both groups regarding the fertilization rate (p = 0.853). There was no statistical difference between them regarding implantation rate (p = 0.237). The percentage of Class A embryos in the ca ionophore group was 81.7%, while it was 82.8% in the control group. There was insignificant difference between them (p = 0.782). There was no statistical significant difference between the two groups regarding the clinical pregnancy rate, it was (56.7%) in the ca ionophore group while it was (53.3%) in the control group. Conclusion: AOA by Ca2++ ionophore didn’t improve the outcome of ICSI cycle in cases of non obstructive azoospermia in terms of fertilization, implantation and pregnancy rate.

Rescue ICSI: Choose the Optimal Rescue Window before Oocyte Aging

Objective To evaluate the application value of rescue ICSI in fertilization failure after conventional IVF and choose the best rescue window before oocyte aging according to the results of rescue ICSI performed in different time. Methods The data of 93 IVF cycles were analyzed retrospectively. Rescue ICSI was performed in these cycles after conventional IVF failure. Because of the different rescue time, these cycles were divided into two groups： early rescue group （group A, 77 cycles, rescue ICSI performed 4-8 h after conventional IVF） and late rescue group （group B, 16 cycles, rescue ICSI performed 20-22 h after conventional IVF）. Results There were no statistically significant differences in age of female, duration of infertility, number of oocytes retrieved every cycle. The normal fertilization rate, pregnancy rate and implantation rate were decreased in group B compared with those in group A （P〈0.05）. In group A, the normal fertilization rate of rescue ICSI performed 4-6 h after conventional IVF （group A1） was increased compared with that of rescue ICSI performed 6-8 h （including 6 h） after conventional IVF （group A2）（66.5% vs 55.9%）（P〈0.05）; while the abnormal fertilization rate in group A1 was decreased compared with group A2 （9.0% vs 14.4%）（P〈0.05）. Clinical pregnancy rate was slight higher in group A1 than in group A2, though this failed to be significantly different. Conclusion Rescue ICSI is effective if fertilization was failure after conventional IVF, the most important thing is to choose the reasonable rescue window before oocyte aging when ICSI is performed.

Immunofluorescent Study of Human MⅡ Oocytes Failed to Fertilize after IVF and ICSI

Objective To discuss the reason why human M Ⅱ oocytes failed to fertilize after IVF and ICSI. Methods The unfertilized human MⅡ oocytes were collected 24-48 h after IVF and ICSI and stained for immunoflurescence and PI counterstain. The types of fertilization failure were identified under the fluorescence microscopy. Results About 55.8% oocytes in IVF were found no sperm in them, which were more than that in ICSI （9.7%） （P〈0.01）. About 14.9% oocytes in IVF and 58.1% in ICSI displayed oocyte activation failure. The difference was significant （P〈0.01）. Defects in pronuclear formation and or migration was found in a similar proportion of oocytes both after IVF （25.3%） and ICSI （32.3%）（P〉0.05）. There were 3.9% oocytes with other abnormalities were observed in IVF but none in ICSI. Conclusion The main reason of fertilization failure after IVF was no sperm penetration. However fertilization failure after ICSI was mainly associated with incomplete oocyte activation.