In vitro maturation of human oocytes maintaining good development potential for rescue intracytoplasmic sperm injection with fresh sperm

BACKGROUND The outcomes of the use of commercial in vitro maturation(IVM)medium to culture immature oocytes obtained from conventional ovulation induction,followed by rescue intracytoplasmic sperm injection(RICSI),are not ideal.It is thus difficult to widely adopt this approach in clinical practice.Therefore,it is necessary to explore methods for improving the clinical outcome of IVM.AIM To study the effect of sperm on the developmental potential of in vitro-matured oocytes in conventional culture.METHODS This was a retrospective study of patients whose immature oocytes were harvested from conventional oocyte stimulation cycles and underwent ICSI at our hospital between June 2018 and August 2020.RICSI was performed using sperm collected on the day of oocyte harvest(old)and sperm collected on the day of RICSI(fresh)and oocytes matured in vitro after 24 h of culture in conventional medium.The rates of in vitro oocyte maturation,normal fertilization,normal cleavage,day-3 top-quality embryos,and useful blastocyst formation were compared between the two groups.RESULTS In total,102 germinal vesicle(GV)-stage immature oocytes were cultured in the old sperm group.In the fresh sperm group,122 GV-stage immature oocytes were collected and cultured in vitro for 24 h.There were no significant differences in the general conditions of males and females between the two groups(P>0.05).The oocyte maturation,normal fertilization,and normal cleavage rates of the old and fresh groups were 51.0%vs 55.7%,61.5%vs 64.7%,and 93.8%vs 93.2%,respectively.None of the rates differed significantly(P>0.05)between the two groups.However,the day-3 top-quality embryo and useful blastocyst rates of the old and fresh sperm groups were 16.6%vs 63.4%;6.67%vs 34.6%,respectively.The day-3 top-quality embryos and useful blastocyst rates of the old sperm group were significantly lower than those of the fresh group(P<0.05).CONCLUSION In vitro maturation with conventional culture medium combined with the use of fresh sperm collected on the day of RICSI is an easy-to-implement strategy for patients whose oocytes are completely or mostly immature.

Genetic and epigenetic risks of intracytoplasmic sperm injection method

Pregnancies achieved by assisted reproduction technologies, particularly by intracytoplasmic sperm injection （ICSI） procedures, are susceptible to genetic risks inherent to the male population treated with ICSI and additional risks inherent to this innovative procedure. The documented, as well as the theoretical, risks are discussed in the present review study. These risks mainly represent thatconsequences of the genetic abnormalities underlying male subfertility （or infertility） and might become stimulators for the development of novel approaches and applications in the treatment of infertility. In addition, risks with a polygenic background appearing at birth as congenital anomalies and other theoretical or stochastic risks are discussed. Recent data suggest that assisted reproductive technology might also affect epigenetic characteristics of the male gamete, the female gamete, or might have an impact on early embryogenesis. It might be also associated with an increased risk for genomic imprinting abnormalities.

Clinical pregnancies and livebirths achieved by intracytoplasmic injection of round headed acrosomeless spermatozoa with and without oocyte activation in familial globozoospermia： case report

We report the successful outcome of intracytoplasmic sperm injection （ICSI） treatment in two siblings with familial globozoospermia. After controlled ovarian hyperstimulation and oocyte pick-up, retrieved oocytes were mechanically activated before ICSI and a fertilization rate of 33.3% was achieved in the first case. The second couple underwent ICSI without oocyte activation and a 9.1% fertilization rate was obtained. The transfer of two grade I embryos in the first couple and one grade I embryo in the second couple resulted in clinical pregnancies with healthy livebirths. It was concluded that the main problem of cases with globozoospermia is a low fertilization rate, and even though ICSI and oocyte activation can increase this rate it is not necessarily needed to achieve a pregnancy. （Asian J Androl 2008 Mar; 10： 332-336）

Pregnancies established through intracytoplasmic sperm injection (ICSI) using spermatozoa with dysplasia of fibrous sheath

Aim:Dysplasia of the fibrous sheath(DFS)is an anomaly found in asthenozoospermic patients with extremely lowor absent motility.In order to determine the efficacy of ICSI in these patients,a retrospective analysis of ICSI results inDFS patients has been done.Methods:Ten ICSI attempts were performed in 6 patients with diagnosis of Dysplasiaof the Fibrous Sheath studied by transmission and scanning electron microscopy.Results:In the cases studied,sperm concentration was(29.62±18.05)×10^(6)/mL,total motility was 1.14±1.31%.Progressive motility was 0%except for one case with 0.1%.One hundred and three preovulatory oocytes were obtained and 94 metaphaseⅡoocyteswere injected.Sixty-nine of them showed two pronuclei(fertilization rate:73.4%).Forty-nine embryos were ob-tained and 34 were transferred(mean:3.4 embryos per transfer).Five pregnancies were diagnosed byβ-hCG plasmalevel determinations that resulted to be one preclinical abortion,one clinical abortion and three deliveries.Anotherpregnancy(ongoing)was achieved from a cryopreserved embryo transfer.Conclusion:These results showed thatICSI provides a suitable solution for patients suffering from irreversible sperm defects such as DFS.Nevertheless,it ismandatory to inform couples of possible transmission risks to offspring,which are unknown at present.Only when theetiology of this problem is disclosed,it will be possible to assess the real genetic risk.

Fertilization of in vitro matured human oocytes by intracytoplasmic sperm injection (ICSI) using ejaculated and testicular spermatozoa

Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.

A nomogram predicting clinical pregnancy in the first fresh embryo transfer for women undergoing in vitro fertilization and intracytoplasmic sperm injection(IVF/ICSI) treatments

The extent to which factors affect the probability of clinical pregnancy in the first fresh embryo transfer after assisted conception is unknown.In order to examine the predictors of clinical pregnancy,a retrospective cohort study was launched between January 1,2013 and December 31,2016 in four infertility clinics including 19837 in vitro fertilization and intracytoplasmic sperm injection(IVF/ICSI)fresh cycles with known outcomes and relevant records.A multivariable logistic regression was used to select the most significant predictors in the final nomogram for predicting clinical pregnancy.Furthermore,the model was validated by an independent validation set and the performance of the model was evaluated by the receiver operating characteristic(ROC)curves along with the area under the ROC curve(AUC)and calibration plots.In a training set including 17854 participants,we identified that female age,tubal factor,number of embryos transferred,endometrial thickness and number of good-quality embryos were independent predictors for clinical pregnancy.We developed a nomogram using these five factors and the predictive ability was 0.66 for AUC(95%CI=0.64−0.68),which was independently validated in the validation set(AUC=0.66,95%CI=0.65−0.68).Our results show that some specific factors can be used to provide infertile couples with an accurate assessment of clinical pregnancy following assisted conception and facilitate to guide couples and clinicians.

Birth after intracytoplasmic sperm injection of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome

Aim:To report a birth after intracytoplasmic sperm injection (ICSI) of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome detected by fluorescence in situ hybridization (FISH) analysis.Methods:A 35-year- old man with a normal appearance consulted our hospital because of sterility over a 5-year period.Chromosome analysis showed low-incidence mosaic Klinefelter's syndrome.Using FISH,96 % hyperploidy of the lymphocytes was found.We examined the sex chromosome of the ejaculated spermatozoa.Using FISH,we examined 200 ejacu- lated spermatozoa and no hyperploidy was found.Results:The 33-year-old female partner of the male patient underwent an uncomplicated controlled ovarian hyperstimulation sequence using a combined recombinant-follicle stimulating hormone (rec-FSH) + human menopausal gonadotrophin (hMG) protocol,following late luteal phase pituitary down regulation.This culminated in the retrieval of seven oocytes,six of which were fertilized with ICSI. One ICSI attempt led to clinical pregnancy with a healthy baby girl.Conclusion:We report a male patient with low- incidence mosaic Klinefelter's syndrome whose ejaculated spermatozoa were identified as being haploid by FISH before ICSI,leading to the successful pregnancy of his wife and the birth of a healthy baby girl.

Effect of Vitamin D Supplementation on Intracytoplasmic Sperm Injection Outcomes: A Randomized Double-Blinded Placebo-Controlled Trial

OBJECTIVE: To assess the effect of vitamin D supplementation on enhancement of embryos implantation rates in intracytoplasmic sperm injection procedures. STUDY DESIGN: A randomized double-blinded controlled trial. SETTING: Assisted Reproduction Unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. DURATION: From December 1st 2017 to the end of October 2018. SUBJECTS and METHDS: Eighty infertile couples that underwent ICSI procedure were entrapped in this study. Patients were divided into two groups;Group I (Vitamin D group) = 40 patients (received vitamin D supplementation in the form of Vidrop oral drops 600 IU/day starting after ovum pickup) and Group II (placebo group) = 40 patients (received a placebo “normal saline oral drop”). RESULTS: There was a significantly increased embryo implantation rates (53.2% in vitamin D group versus 46.7% in placebo group), there was a mild statistically significant difference in the clinical pregnancy rates (45.9% in vitamin D group versus 39.4% in placebo group) with p-value < 0.05 and there was an improvement in ongoing pregnancy rate as there was a mild statistically significant difference in first trimester miscarriage rates (12.5% in vitamin D group versus 17.5% in placebo group) with p value CONCLUSIONS: Vitamin D supplementation could improve embryos implantation rates and rate of ongoing pregnancy by decline in the first trimester miscarriage rate.

Role of Dehydroepiandrosterone Supplementation in Improving Intracytoplasmic Sperm Injection Outcome for Women with Expected Poor Ovarian Response

Background: As regard to adjuvant supplementations, nowadays dehydroepiandrosterone (DHEA) is widely used all over the world and is considered to be a potential agent to ameliorate the assisted reproduction technologies outcomes of infertile women with poor ovarian reserve. Objective: To find out the role of DHEA supplementation in improving intracytoplasmic sperm injection (ICSI) outcome for infertile women with expected poor ovarian response in controlled ovarian stimulation. Setting: Assisted reproduction unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. Duration: From April 2016 to May 2018. Study Design: A randomized double-blinded controlled trial. Methods: One hundred and forty infertile women with expected poor ovarian response prepared for ICSI procedure were included in this study. Patients were divided into two groups;group I (DHEA group) included 70 patients received 25 mg DHEA 12 weeks prior to ICSI cycle and group II (placebo group) included 70 patients received a placebo. Results: There was a highly statistically significant difference in basal AFC at start of ICSI cycle in group I (who received DHEA supplementation for 12 weeks prior to ICSI procedure) than in group II (13.8 ± 5.3 versus 10.7 ± 4.6 respectively) with P < 0.001. There were mildly statistically significant differences between group I and group II as regard to increase in the number and quality of retrieved oocytes, increased in endometrial thickness, fertilization rate and embryo quality with p value < 0.05 but there was no statistically significant difference between the 2 groups as regard to pregnancy (chemical and clinical) rates (p value > 0.05). Conclusions & Recommendations: DHEA supplementations improved basal AFC, increased the number & quality of oocytes and increased quality of embryos in infertile patients with expected poor ovarian response in ICSI procedure. So DHEA supplementations could be an important adjuvant for infertile women with expected poor ovarian response in ICSI procedure.

Use of hyaluronic acid for sperm immobilisation and selection before intracytoplasmic sperm injection:A systematic review and meta-analysis

AIM:To appraise critically the published randomised controlled trials(RCTs)reporting on the effectiveness of using hyaluronic acid(HA)for sperm immobilisation and selection before intracytoplasmic sperm injection(ICSI).METHODS:Two authors used the PICO Method in order to perform a comprehensive literature search of the standard medical databases in June 2015.Data from the included studies was extracted independently by two authors using a predefined pro-forma.Review Manager(RevM an)was used to calculate the combined outcomes where multiple studies contributed with their results.Risk ratio(RR)with a 95%CI using the Mantel-Haenszel method was calculated for binary data variables.Heterogeneity was measured using the χ2 test and quantified using I2.In case of substantial heterogeneity(P < 0.10 for χ2 test or I2 > 50%)the combined outcome was calculated using the random effects model.The results from the meta-analysis were displayed as forest plots.The guideline of the Cochrane Collaboration was used to assess the risk of bias and it was illustrated as a risk of bias graph.RESULTS:The systematic literature search identified 166 different studies related to sperm immobilisation and selection for ICSI.Eleven RCTs involving 13719 oocyte intracytoplasmatic injections with sperm immobilised and selected using HA or polyvinylpyrrolidone(PVP)were included in this systematic review and metaanalysis.There was low heterogeneity among the included trials(χ2 = 16.86,df = 11,P = 0.11; I2 = 35%).There was no statistical difference between HA and PVP groups in terms of fertilisation rate(RR = 1.01; 95%CI:0.99-1.03; z = 0.75; P = 0.45),good embryos rate(RR = 1.01; 95%CI:0.96-1.06; z = 0.30; P = 0.76),live birth rate(RR = 1.15; 95%CI:0.86-1.54; z = 0.92; P = 0.36),clinical pregnancy rate(RR = 1.04; 95%CI:0.92-1.17; z = 0.62; P = 0.53)and implantation rate(RR = 1.17; 95%CI:0.94-1.46; z = 0.40; P = 0.16).The quality of most of the included studies was moderate to poor because of unclear randomisation technique,inadequate allocation concealment and blinding.CONCLUSION:This systematic review and metaanalysis provides evidence of similar efficiency between using HA or PVP for sperm immobilisation and selection before ICSI.

Patients with MMAF induced by novel biallelic CFAP43 mutations have good fertility outcomes after intracytoplasmic sperm injection

As a specific type of asthenoteratozoospermia,multiple morphological abnormalities of the sperm flagella(MMAF)is characterized by composite abnormalities,including absent,short,coiled,angulation,and irregular-caliber flagella.Mutations in cilia-and flagella-associated protein 43(CFAP43)are one of the main causative factors of MMAF established to date.To identify whether there are other CFAP43 mutations related to MMAF and to determine the clinical outcomes of assisted reproductive technology for patients with MMAF harboring different mutations,we recruited and screened 30 MMAF-affected Chinese men using a 22-gene next-generation sequencing panel.After systematic analysis,seven mutations in CFAP43,including five novel mutations and two previously reported mutations,were identified from four families and related to MMAF in an autosomal recessive pattern.Papanicolaou staining,immunofluorescence,and electronic microscopy further clarified the semen characteristics a nd abnormal sperm morphologies,including disorganized axonemal and peri-axonemal structures,of the CFAP43-deficient men.The female partners of two patients were pregnant after undergoing assisted reproductive technology through intracytoplasmic sperm injection,and one of them successfully gave birth to a healthy boy.This study significantly expands the mutant spectrum of CFAP43,and together with the available information regarding male infertility and MMAF,provides new information for the genetic diagnosis and counseling of MMAF in the future.

Use of testicular sperm for intracytoplasmic sperm injection in men with high sperm DNA fragmentation： a SWOT analysis

Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation （SDF） in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology （ART）, an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection （Testi-ICSI）. In this article, we used a SWOT （strengths, weaknesses, opportunities, and threats） analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval.

Testicular sperm extraction and intracytoplasmic sperm injection in non obstructive azoospermia

Objective To report the experience in sperm extraction from testicular biopsies (TESE) performed from March 1996 to July 1998 in men with non obstructive azoospermia (NOA) Methods Comparisons of age, volume of both testes, serum FSH and testosterone in men, and histology of testicular samples in the first cycles between cycles with spermatozoa found and those without spermatozoa found were performed Comparisons of fertilization, cleavage and pregnancy rates between cycles with spermatozoa injected and those with spermatids injected were performed Results Spermatozoa were found in only 12 out of 26 first TESE cycles (46 2%) and other cycles had spermatids (round cells) only Age of men, history of mumps orchitis/oligozoospermia, volume of both testes and serum FSH/testosterone levels in men were not significantly different between cycles with and without spermatozoa The fertilization rate was significantly higher in cycles with spermatozoa injected than those with round cell injections (63 3% vs 23 2%, P <0 0001, Chi squared test) The pregnancy rate was 14 3% per cycle when spermatozoa were injected Conclusions TESE followed by Intracytoplasmic sperm injection (ICSI) is an effective treatment in patients with NOA Less than half of the patients undergoing TESE had spermatozoa recovered Age of men, volume of both testes and serum FSH/ testosterone levels in men were not useful in predicting successful recovery Compared to using ejaculated and epididymal spermatozoa, fertilization and pregnancy rates were achieved when testicular spermatozoa were used for ICSI

Different sperm sources and parameters can influence intracytoplasmic sperm injection outcomes before embryo implantation

To evaluate the effects of sperm with different parameters and sources on the outcomes of intracytoplasmic sperm injection (ICSI), 1972 ICSI cycles were analyzed retrospectively. Groups 1 to 5 were composed of cycles using ejaculated sperm and were grouped according to sperm quantity, quality, and morphology into normal (288 cycles), or mild (329 cycles), moderate (522 cycles), severe (332 cycles), and extremely severe (171 cycles) oligozoospermia and/or asthenozoospermia and/or teratozoospermia (OAT) groups. Group 6 was composed of 250 cycles using testicular or epididymal sperm, and Group 7 consisted of 80 cycles using frozen-thawed sperm. We found that fertilization rates were gradually reduced from Groups 1 to 6, and reached statistical difference in Groups 5 and 6 (P<0.05). The high-quality embryo rate was higher in Group 1 than in Groups 2, 3, 5, 6, and 7 (P<0.05). No statistical differences were observed in the rates of embryo cleavage, clinical pregnancy, miscarriage, live-birth, premature birth, low birth weight, weeks of premature birth, average birth weight, or sex ratio for all seven groups (P>0.05). A total of nine cases of malformation were observed, with a malformation rate of 1.25% (9/719). In conclusion, different sperm sources and parameters can affect ICSI outcomes before embryo implantation. A full assessment of offspring mal-formation will require further study using a larger sample size.

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.

Follow-up of children born after intracytoplasmic sperm injection with epididymal and testicular spermatozoa

Background To evaluate the safety of intracytoplasmic sperm injection （ICSI） with epididymal or testicular sperm, this study compared children born after ICSI treatment with epididymal or testicular sperm with children conceived after ICSI with ejaculated sperm. Methods This retrospective study included 317 children born after ICSI with percutaneous epididymal sperm aspiration （PESA）, 103 children born after ICSI with testicular sperm aspiration （TESA）, and a control group of 1008 children born after ICSI with ejaculated sperm. All of the patients received their assisted reproductive treatment in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2004 to December 2011. Data, such as the rate of stillbirths, perinatal mortality, gestational age, birth weight, and the rate of congenital malformations of the three groups, were compared. Results PESA and TESA children were not different from ICSI children in the rate of stillbirths, perinatal mortality, infant mortality rate, gestational age, the rate of prematurity, and the rate of malformations （P〉0.05）. A slight increase in birth defects was reported in the TESA group compared with those in the control group, but there was no significant difference between the groups （P〉0.05）. Conclusion ICSI with epididymal or testicutar sperm does not lead to more stillbirths or congenital malformations compared with ICSI using ejaculated sperm.

Success rates of in vitro fertilization versus intracytoplasmic sperm injection in men with serum anti-sperm antibodies:a consecutive cohort study

Antisperm antibodies(ASAs)are assumed to be a possible causative factor for male infertility,with ASAs detected in 5%-15%of infertile men but in only l%-2%of fertile ones.It remains unclear whether ASAs have an adverse effect on the outcome of in v/tro fertilization(IVF)or intracytoplasmic sperm injection(ICSI).This study investigated differences in the rates of fertilization,pregnancy,and live births associated with serum ASA-positive and ASA-negative men following IVF or ICSI.Five hundred and fifty-four consecutive infertile couples undergoing IVF(n=399)or ICSI(n=155)were included.The two-sample two-sided t-test and Chi-square or Fisher's exact test was used for statistical analysis.Lower rates of fertilization(41.7%vs 54.8%,P=0.03),good embryos(18.9%vs 35.2%,P=0.00),pregnancy(38.5%vs 59.4%,P=0.00),and live births(25.8%vs 42.5%,P=0.00)were observed in men of the IVF group with a positive serum ASA than in those with a negative ASA.ASA positivity/negativity correlated with pregnancy rates(P=0.021,odds ratio[OR]:0.630,95%confidence interval[Cl]:0.425-0.932)and live birth rates(P=0.010,OR:1.409,95%Cl:1.084-1.831)after controlling for the female serum follicle-stimulating hormone level and the couple's ages at IVF.Women coupled with ASA-positive men had lower live birth rates with IVF than with ICSI(25.8%and 47.4%,respectively;P=0.07).Women coupled with ASA-positive men had lower rates of pregnancy and live births following IVF than those coupled with ASA-negative men but had a similar outcome with ICSI.

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).

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.

Should few retrieved oocytes be as an indication for intracytoplasmic sperm injection?

Objective:To reevaluate whether relatively few oocytes obtained in one cycle are an indication for intracytoplasmic sperm injection(ICSI).Methods:A total of 406 cycles with three or fewer retrieved oocytes performed in 396 non-male infertile couples were retrospectively reviewed.Cycles were classified into three groups by different fertilization techniques:the in vitro fertilization(IVF) group,insemination with conventional IVF;the ICSI group,insemination with ICSI though semen parameters were normal;and the rescue ICSI group,re-insemination with ICSI after conventional IVF failure.Results:The ICSI group resulted in higher normal fertilization compared with the conventional IVF group.Correspondingly,the cycle cancellation rate was decreased in the ICSI group,though it was not statistically significant.The clinical pregnancy rate and implantation rate were lower in the ICSI group compared with the conventional IVF group.Rescue ICSI was a method to avert total fertilization failure in conventional IVF,increasing fertilization and ensuring embryo availability for transfer,but the normal fertilization was the lowest due to delayed insemination and the chance of pregnancy was very little.Conclusions:Obtaining only few oocytes in one cycle is not considered as an indication for ICSI when the sperm sample is apparently normal.Rescue ICSI is either not recommended if conventional insemination fails.Such patients should not be subjected to the unnecessary costs and potential risks of ICSI.