Fresh Intracytoplasmic Sperm Injection Research Articles

Birth of Kittens After the Transfer of Frozen–Thawed Embryos Produced by Intracytoplasmic Sperm Injection with Spermatozoa Collected from Cryopreserved Testicular Tissue

The aim of this study is to produce live kittens from oocytes fertilized by intracytoplasmic sperm injection (ICSI) with frozen/thawed testicular spermatozoa. Spermatozoa were collected from thawed testicular tissue and subsequently injected into in vitro matured cat oocytes. At 24 h post-ICSI, presumptive zygotes/cleaved embryos were treated with 10 μm forskolin for 24 h to reduce intracellular lipid content of embryos (delipidation). At 48 h after oocyte injection, cleaved embryos (2- to 8-cell stage) were frozen in 10% (v/v) ethylene glycol-based medium by a slow controlled rate method and stored in liquid nitrogen. To evaluate in vitro and in vivo developmental competence, frozen embryos were thawed and then cultured for 6 days (n = 155) or cultured for 2 h before transferred (n = 209) to hormonal (equine chorionic gonadotropin/hCG)-treated cat recipients. Cleavage frequency at day 2 after ICSI with frozen/thawed testicular spermatozoa was ~30%. The percentages of frozen/thawed embryos that developed to morula and blastocyst stage (on day 3 and day 6 of in vitro culture, respectively) were significantly lower than that of fresh ICSI embryos (22.6 vs 45.2% and 21.3 vs 38.7%, respectively; p < 0.05). However, no difference was found in the number of blastomeres between frozen/thawed (242.5 ± 43.1) and fresh (320.2 ± 28.1) blastocysts. Three of seven cat recipients were pregnant and one pregnant cat delivered two healthy kittens. This is the first report of the birth of kittens after the transfer of frozen-thawed embryos produced by ICSI with frozen/thawed testicular sperm.

Management of endometrial polyps diagnosed before or during ICSI cycles

This retrospective study aimed to shed light on the management options of endometrial polyps diagnosed before or during intracytoplasmic sperm injection (ICSI) treatment. The study included all fresh ICSI cycles performed in the Anatolia IVF Center between July 2005 and January 2009. Group 1 consisted of 47 patients who were diagnosed with an endometrial polyp before their ICSI cycle. All patients diagnosed with an endometrial polyp by transvaginal ultrasonography before the ICSI cycle underwent hysteroscopic polyp resection. Group 1 was compared with 47 matched control patients without endometrial polyps who underwent standard ICSI cycles (group 2). Group 3 included 128 patients diagnosed with an endometrial polyp during stimulation in their ICSI cycles. Group 3 was compared with 128 matched control patients without endometrial polyps who underwent standard ICSI cycles (group 4). Patients diagnosed with an endometrial polyp before ICSI cycles were similar to their controls with regard to clinical pregnancy (29.8% versus 38.3%) and live-birth (25.5% versus 31.9%) rates per transfer, as were patients diagnosed with an endometrial polyp during ovarian stimulation (clinical pregnancy rates 45.3% versus 46.9%; live-birth rates 40.6% versus 39.8%). In conclusion, further studies are required to identify the most appropriate management of endometrial polyps. In our retrospective study, we aimed to shed light on management options of endometrial polyps diagnosed before or during intracytoplasmic sperm injection (ICSI) treatment. This study included all fresh ICSI cycles performed at the Anatolia IVF Center between July 2005 and January 2009. Group 1 consisted of 47 patients who were diagnosed with an endometrial polyp before their ICSI cycle. All patients diagnosed with an endometrial polyp by transvaginal ultrasonography before the ICSI cycle underwent hysteroscopic polyp resection. Group 1 was compared was compared to 47 matched control patients without endometrial polyps who underwent standard ICSI cycles (group 2). Group 3 included 128 patients diagnosed with an endometrial polyp during stimulation in their ICSI cycles. Group 3 was compared to 128 matched control patients without endometrial polyps who underwent standard ICSI cycles (group 4). Patients diagnosed with an endometrial polyp before ICSI cycles were similar to their controls with regard to pregnancy rates and outcomes. Patients diagnosed with an endometrial polyp during ovarian stimulation were similar to their controls with regard to pregnancy rates and outcomes. In conclusion, endometrial polyps <1.5 cm occurring during ovarian stimulation do not require any intervention. Resection of an endometrial polyp <1.5 cm detected before an ICSI cycle is not required and might be even harmful.

Frozen-thawed blastocyst transfer reduces ectopic pregnancy risk: an analysis of single embryo transfer cycles in Japan

To evaluate the effect of frozen-thawed blastocyst transfer on ectopic pregnancy (EP) rate. Retrospective analysis. Japanese registry of assisted reproductive medicine. Women who were registered during 2008. None. Rates of live birth, EP, and spontaneous abortion in single blastocyst and cleavage-stage ET cycles in both fresh and frozen-thawed were determined. In the frozen-thawed single blastocyst transfer cycles, 84 EPs of 10,312 clinical pregnancies (0.81%) were reported. There were 25 EPs of 1,361 clinical pregnancies (1.8%) in fresh IVF single blastocyst transfer cycles and 19 of 1,352 (1.4%) in fresh intracytoplasmic sperm injection single blastocyst transfer cycles. The rate of EPs after frozen-thawed single blastocyst transfers varied between 0.56% and 0.97% after stratification by age group. Frozen-thawed blastocyst transfers have reduced EP rates significantly compared with fresh blastocyst transfers.

A 10-year survey on birth defects after in vitro fertilization-embryo transfer in Shanghai

To survey birth defects of neonates conceived by using various types of in vitro fertilization and embryo transfer (IVF-ET) between 1998 and 2007 in Shanghai. From 1998 to 2007, 8507 neonates from 6551 pregnancies conceived through assistant reproductive technology (ART) from 7 reproductive medicine center in Shanghai were enrolled in this retrospective study, including Shanghai Ji-Ai Genetics and IVF Institute, Shanghai Jiaotong University School of Medicine affiliated Renji Hospital, Ruijin Hospital, China Welfare Institute International Maternal and Infant Health Hospital, Shanghai First Maternity and Infant Health Hospital, Shanghai the Ninth People's Hospital and the Second Military Medical University affiliated Changhai Hospital. The clinical data about the type and incidence of birth defect were analyzed. Meanwhile, the factors associated with birth defect were investigated, such as various ART, gender, mother age, numbers of gestational sac, the source and quality of sperm. The birth defect rate was 1.08% (92/8507) and the most remarkable malformation occurred in circulation system [34% (31/92)]. The rates of major congenital malformations were 1.21% (34/2799) in fresh conventional IVF-ET, 1.07% (20/1871) in IVF-frozen-thawed embryo transfer (IVF-FET), 1.04% (23/2212) in fresh intracyto plasmic sperm injection (ICSI)-ET and 0.92% (15/1625) in ICSI-FET, which did not show statistical difference (P > 0.05). There was also no significantly different incidence of malformations between 1.12% (49/4371) in male neonates and 1.02% (42/4136) in female neonates (P > 0.05). However, the occurrence of congenital malformation is positively related with maternal age, the rates were 0.84%(41/4884) in mother's age less than 30 years and 1.77% (16/902) in mother' age more than 35 years, which reached statistical difference (P < 0.05). It also showed remarkable different incidence among 0.53% (25/4679) in singletons, 1.59% (57/3576) in twins and 3.97% (10/252) in triplets (P < 0.05). No remarkable difference of malformation rate among sperm sources used in fertilization were observed, including 1.09% (81/7419) in ejaculated sperm, 1.21% (7/579) in percutaneous epididymal aspiration (PESA) and 0.79% (4/509) in donor sperm (P > 0.05). The overall incidence of birth defect in neonates from ART is similar to those conceived naturally and is not associated with different types of IVF-ET, embryo cryopreservation, sperm sources. However, maternal age and multiple pregnancies confer the higher possibility of birth defect.

Poor sperm quality negatively impacts embryo development and reproductive outcome in donor oocyte assisted reproductive technology cycles

OBJECTIVE: Oocyte donation offers a powerful tool in the study of sperm quality, as oocyte and endometrial characteristics can be controlled and are comparable among the recipients. Since the introduction of intracytoplasmic sperm injection (ICSI), little attention is given to specific parameters of sperm function and their impact on IVF outcome. Our aim is to determine whether semen parameters influences embryo development and reproductive potential, and if they could be regarded as a prognostic factor in donor assisted reproductive technology cycles. DESIGN: Retrospective cohort analysis. MATERIALS AND METHODS: Three hundred and thirty two fresh ICSI cycles using donor oocytes were analyzed. ICSI was performed routinely to avoid inmature oocytes being given to the recipients. Only cycles in which >3 embryos were yielded and at least two Day3 good quality embryos were available for ET were included. Male partners were classified as normal (N), low-moderate (LM) or severe (S) male factor infertility according to the following sperm parameters: 1) Percent normal morphology by strict Kruger criteria: (S) <4%, (LM) 4-14%, (N)>14%, 2) Sperm concentration: (S) <10million/ml, (LM) 10- 20 million/ml, and (N) > 20million/ml, 3) Motility (L) <30%, (LM) 30-50%, (N) >50%, and 4) Post-preparation total motile count: (S) < 5 million, (LM) 5-10 million, and (N)>10 million sperm. Male factor infertility was correlated to a variety of outcome measures: embryo quality (day-3 cell cleavage, and fragmentation), fertilization, clinical pregnancy rate (CPR) and pregnancy loss. Statistical analysis was performed with chi2 for categorical data and t test for continuous parametric data. RESULTS: The number of fertilized oocytes and the number of day 3 embryos >7 cells was similar among the normal and low-moderate factor infertility group. However, the number of good quality embryos available for ET were significantly lower in the severe male factor infertility group (P<0.005). Implantation and clinical pregnancy rates were significantly decreased in this group when compared to male partners with normal or low-moderate semen parameters: Normal CPR 52,05% (76/146), Low-Moderate CPR 40,91%, Severe CPR 29,63% (16/54) (p<0.05). CONCLUSIONS: Poor sperm quality was negatively associated with reproductive outcomes in oocyte donation cycles suggesting a paternal influence on embryo development in which ICSI may not be able to select the sperm of highest quality for oocyte injection.

Poor sperm quality affects clinical outcomes of intracytoplasmic sperm injection in fresh and subsequent frozen–thawed cycles: potential paternal effects on pregnancy outcomes

To evaluate objectively whether poor sperm quality affects sequential events from fertilization to delivery in fresh intracytoplasmic sperm injection (ICSI) and subsequent frozen-thawed embryo transfer (ET) cycles. Retrospective study. University-based centers for reproductive medicine. For unbiased comparison, 206 cycles were chosen from 1,999 cycles of patients who underwent ICSI-ET and/or subsequent frozen-thawed ET. Cycles met the following criteria: day 3 ET; female age, <40 y; number of retrieved oocytes, >or=5; no split insemination; and no female factors but tubal factor. None. The rates of fertilization, embryo implantation, clinical pregnancy, and delivery and sequential embryonic score (SES) were compared between normal-spermatogenesis patients (NSPs) and defective-spermatogenesis patients (DSPs). Although sum SES, mean SES, and top SES of transferred embryos on day 3 were similar between NSPs and DSPs, the rates of implantation, clinical pregnancy, and delivery of NSPs were significantly higher than those of DSPs. Furthermore, subsequent ET cycles with frozen-thawed embryos in NSPs and DSPs who failed to achieve pregnancy in their fresh cycles showed that rates of implantation and clinical pregnancy also were significantly lower in DSPs. Quality of sperm may influence embryo implantation and subsequent pregnancy outcomes without impairment of embryo quality.

Progesterone levels on oocyte retrieval day can predict the quantity of viable embryos but not pregnancy outcome of intracytoplasmic sperm injection

The present study aimed to determine whether serum progesterone (P) concentration on the day of oocyte retrieval (OR) affects the quantity of viable embryos and impacts the outcome of intracytoplasmic sperm injection (ICSI). A total of 289 women who underwent ICSI cycles after controlled ovarian hyperstimulation were divided in two groups based on P level (<11.7 and ≥ 11.7 ng/ml) on OR day. Subjects did not differ significantly with respect to age, basal follicle-stimulating hormone levels, body mass index, or estradiol levels on day of administration of human chorionic gonadotropin or OR day. Women with ICSI cycles that had high P levels tended to have more retrieved oocytes, zygotes and embryos than women with lower levels, but the differences did not approach significance. However, the former group with higher P levels did have a significantly greater number of viable embryos (p = 0.003). Finally, the two groups did not show differences in pregnancy outcome in a new ICSI cycle. The study suggests that serum P levels on OR day are related to the quality of viable embryos but not to pregnancy outcome in fresh ICSI cycles.

South Asian women with polycystic ovary syndrome exhibit greater sensitivity to gonadotropin stimulation with reduced fertilization and ongoing pregnancy rates than their Caucasian counterparts

Polycystic ovary syndrome (PCOS) is a heterogeneous syndrome. In vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) is required for PCOS cases that are refractory to standard ovulation induction or have co-existing infertility factors in women with PCOS and Tubal factor subfertility. Objectives Assess ethnic variations in response to IVF/ICSI treatment. Study design Observational Comparative study in a University hospital fertility clinic in women with PCOS and Tubal factor subfertility. Women with PCOS (Asians: AP = 104; Caucasians: CP = 220) and those with tubal factor infertility seeking fertility treatment were assessed (Asians: AC = 84; Caucasians: CC = 200). Six hundred and eight fresh IVF or ICSI cycles using long protocol of GnRHa suppression and resulting in a fresh embryo transfer were compared. The primary endpoint was to assess the dose of gonadotropins used in the cycles. The secondary outcomes were: total number of oocytes retrieved, fertilization and ongoing clinical pregnancy rates. Results We found that the South Asian women presented at a younger age for the management of sub-fertility. An extended stimulation phase and Caucasian ethnicity showed an inverse correlation with the number of oocytes retrieved in the PCOS subgroup. Caucasian ethnicity was associated with a higher fertilization rate however increase in body mass index (BMI) and the laboratory technique of IVF appeared to have a negative impact on fertilization rates in the PCOS subgroup. Commencing down regulation on day 1 of the cycles was negatively associated with fertilization rates in the tubal group. In terms of clinical pregnancy rates, the Caucasian PCOS had a 2.5 times (95% CI: 1.25–5) higher chance of an ongoing clinical pregnancy as compared with their Asian counterpart. Also, a unit increase in the basal FSH concentration reduced the odds of pregnancy by 18.6% (95% CI: 1.8–32.6%) in the PCOS group. Conclusions The Asian PCOS have a greater sensitivity to gonadotropin stimulation with lower fertilization and ongoing clinical pregnancy rates as compared with their Caucasian counterparts.

P-719: The effect of smoking on performance of intracytoplasmic sperm injection cycles without another factor threatening ovarian reserve

P-719: The effect of smoking on performance of intracytoplasmic sperm injection cycles without another factor threatening ovarian reserve

P-488: The number of oocyte pick up procedure per a single day may have detrimental effect on fertilization rate in intracytoplasmic sperm injection cycles

P-488: The number of oocyte pick up procedure per a single day may have detrimental effect on fertilization rate in intracytoplasmic sperm injection cycles

Optimal use of fresh and frozen-thawed testicular sperm for intracytoplasmic sperm injection in azoospermic patients

To optimize the use of fresh and frozen-thawed testicular biopsy specimens from patients with azoospermia. Fifty-one patients suffering from obstructive and non-obstructive azoospermia underwent testicular sperm extraction (TESE). The specimens were divided and used for either in vitro maturation or freezing for a future intracytoplasmic sperm injection (ICSI) cycle. At initial testicular sperm extraction, very few motile spermatozoa were seen. After 24 h of in vitro maturation, sperm motility increased remarkably, with a maximum motility rate seen between 48 and 72 h of culture. Motile spermatozoa were observed up to 120 h in culture. In the 22 fresh ICSI cycles, a total of 294 oocytes were injected using motile sperm and 212 oocytes demonstrated normal 2PN formation (fertilization rate, 72.1%). In 36 frozen-thawed ICSI cycles, a total of 454 oocytes were injected and 302 oocytes became 2PN (66.5%). On day 3, high quality embryos were observed in 54.2% of fresh cycles and 54.1% of frozen cycles (P > 0.05). The clinical pregnancy rate did not show a significant difference between using fresh (59%) and frozen (55.5%) testicular biopsy sperm for ICSI (P > 0.05), but the embryo implantation rates did differ significantly between fresh (29.5%) and frozen-thawed (22.2%) cycles (P < 0.05). A total of 33 healthy babies have been born from 22 women, giving birth after 58 embryo transfer attempts (38%). The freezing and in vitro culturing of testicular biopsy tissue is a very reliable approach for the management of testicular biopsy specimens from azoospermic patients, and offers the possibility of several treatments of IVF/ICSI from a single sample.

Intracytoplasmic sperm injection of frozen-thawed bovine oocytes and subsequent embryo development.

Oocyte cryopreservation and intracytoplasmic sperm injection (ICSI) are advantageous to expand their usefulness in genetic engineering. Oocytes matured for 22 hr were vitrified in droplets of cryoprotectants (3.2 M ethylene glycol (EG), 2.36 M dimethyl sulfoxide (DMSO), 0.6 M sucrose) on copper electron microscope (EM) grids. After being warmed, the oocytes were cultured in IVM medium for an additional 2 hr. Sperm treated with dithiothreitol were utilized for ICSI. Oocytes injected with sperm were activated by combination of ionomycin with cycloheximide (CHX). The ICSI oocytes were compared for the rates of pronuclear formation, development, cell number, and the ratio of ICM to those of fresh ICSI and IVF control. The proportion of 2PN formation was significantly higher in IVF control (Group 1) than those in other treated groups. Among the treated groups a significant lower 2PN formation was observed in IVF-frozen-thawed than in ICSI-fresh and frozen-thawed groups. Cleavage rates in IVF-frozen-thawed and ICSI-frozen-thawed groups were significantly lower than those of IVF control and ICSI-fresh groups. In ICSI groups, the rates of cleavage and blastocyst in fresh oocytes were significantly higher than in frozen-thawed. Development rates into blastocysts in the ICSI-fresh and frozen-thawed groups were significantly lower than that of IVF control. Total cell number was significantly lower in both frozen-thawed IVF and ICSI groups than those in IVF-control and ICSI-fresh groups. However, the rates of the remaining cells that were found in the ICM were significantly higher in both frozen-thawed IVF and ICSI than in the IVF-control and ICSI-fresh groups. The results indicated that frozen-thawed bovine oocytes were suitable for ICSI procedure.

Recombinant or urinary follicle-stimulating hormone? A cost-effectiveness analysis derived by particularizing the number needed to treat from a published meta-analysis

Objective: To demonstrate that particularizing pooled results of a meta-analysis can derive incremental cost effectiveness of superovulation with recombinant follicle-stimulating hormones (rFSH) vs. the highly purified urinary form (uFSH) for assisted conception. Design: A retrospective study. Setting: An assisted conception unit in the United Kingdom. Patient(s): One hundred forty-five fresh in vitro fertilization (IVF) and 58 fresh intracytoplasmic sperm injection (ICSI) cycles. Intervention(s): rFSH vs. uFSH. Main Outcome Measure(s): Incremental cost-effectiveness (i.e., cost needed to treat, or CNT) and budget-impact analyses of rFSH vs. uFSH. Result(s): In women less than 30 years old, the clinical pregnancy rate was 37.7% (95% CI 24.8%–52.1%), the particularized number needed to treat (pNNT) was −19, and the cost needed to treat was £5070.51 (£3660.53 to £7619.32). For the 30- to 35-year-old age group, the clinical pregnancy rate was 29.9% (95% CI 20.0%–41.4%), the particularized number needed to treat was −24, and CNT was £7335.59 (£5284.11 to £10,941.22). For the 36- to 40-year-old age group, the clinical pregnancy rate was 30.6.0% (95% CI 19.6%–43.7%), the particularized number needed to treat was −23.0, and the CNT was £8569.67 (£5998.70 to £13,413.24). Conclusion(s): The CNT and thus the budget impact analyses (the extra number of cycles that can be funded by the CNT) both increase directly with age of the patient, and inversely with the clinical pregnancy rate.

Obstetric outcome of pregnancies after the transfer of cryopreserved and fresh embryos obtained by conventional in-vitro fertilization and intracytoplasmic sperm injection.

This study reports the obstetric outcome of pregnancies obtained after the transfer of cryopreserved or fresh embryos where the initial procedure was standard in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Pregnancies obtained after frozen IVF (n = 245) or frozen ICSI (n = 177) were compared with a control group of pregnancies after fresh embryo transfer in standard IVF (n = 245) and ICSI (n = 177) cycles were selected as controls. The controls were matched according to maternal age, parity and date of embryo transfer. In the standard IVF group, the biochemical pregnancy rates in the cryopreserved and fresh groups were 18.8 and 9.8% respectively (P < 0.01). In the ICSI group, the biochemical pregnancy rates in the cryopreserved and fresh groups were 16.4 and 6.8% respectively (P < 0.01). The miscarriage rates were comparable between the cryopreserved and fresh groups. However, in the frozen ICSI group the miscarriage rate (26.0%) was significantly higher than in the frozen conventional IVF group (13.1%) (P = 0.001). The frequencies of preterm deliveries, infants with very low birthweight and intrauterine deaths were similar in the groups. The low birthweight rates in the frozen IVF (16.1%) and ICSI (12.1%) groups were significantly lower than those in the fresh IVF (32.2%) and ICSI (32.7%) groups (P < 0.001). The major malformation rates in the frozen IVF (2.4%) and ICSI (2.9%) groups were not different from the major malformation rates in the fresh IVF (4.5%) and ICSI (2.4%) groups. In conclusion, the cryopreservation process had no negative impact on the outcome of pregnancies over 20 weeks of gestation. Long-term follow-up studies are needed in order to prove the safety of the freezing-thawing process.