Laser-assisted intracytoplasmic sperm injection in human oocytes

OBJECTIVE: To explore potential applications of a non-contact, 1,480-nm diode laser to intracytoplasmic sperm injection (ICSI) of human oocytes. STUDY DESIGN: Human oocytes obtained from in vitro fertilization (IVF) patients and failed to fertilize 24-48 hours after conventional IVF were used for ICSI along with discarded sperm. A noncontact, 1,480-nm diode laser was employed to immobilize sperm, to open a hole in the zona pellucida (ZP) and to perform ICSI through the hole. After ICSI and its simulation, oocytes were examined for formation of pronuclei, cleavage and normality of the cytoskeleton. RESULTS: The 1,480-nm diode laser permitted fast and easy sperm immobilization and microdrilling of ZP to facilitate microinjection. Of the 78 injected oocytes, 53 (68%) survived the procedure, and 13 (25%) of them formed two pronuclei by 18 hours. Further culture of two fertilized eggs resulted in cleavage up to the eight-cell stage before cease of culture. Four oocytes were fixed after simulation of the procedure without injecting sperm. None of them showed gross abnormalities in cytoskeletal organization. CONCLUSION: A noncontact, 1,480-nm diode laser can be used for the immobilization of sperm and for opening a hole in the ZP to facilitate ICSI, biopsy manipulation toward preimplantation genetic diagnosis and assisted hatching.     

Maturational and developmental competence of immature oocytes retrieved from bovine ovaries at different phases of folliculogenesis

Immature oocytes recovered from bovine ovaries were studied to determine if their maturational and developmental competence is affected by phase of folliculogenesis. Ovaries (a total of 39 pairs) were collected from a local abattoir. Following examination, each pair of ovaries was assigned to one of three groups, according to follicle size and with or without a corpus luteum: (i) early phase (n = 13 pairs): all follicles were or=15 mm in diameter; (iii) luteal phase (n = 13 pairs): all follicles were 相似文献   

Lack of correlation between oocyte-corona-cumulus complex morphology and nuclear maturity of oocytes collected in stimulated cycles for intracytoplasmic sperm injection

OBJECTIVE: To evaluate the usefulness of morphology grading of the oocyte-corona-cumulus complex (OCCC) as a marker of oocyte nuclear maturity, fertilizability, embryo cleavage, and likelihood of pregnancy. DESIGN: Prospective cohort study. SETTING: Academic fertility center. PATIENT(S): Eighty-three infertile couples undergoing IVF-ET/intracytoplasmic sperm injection treatment. INTERVENTION(S): All patients underwent a long stimulation protocol of GnRH agonist therapy followed by hMG administration and transvaginal oocyte recovery. MAIN OUTCOME MEASURE(S): All OCCCs, oocytes, and embryos were assessed. The relation among OCCC morphology and the nuclear maturity of denuded oocytes, the fertilization rate, and embryo development to the cleavage stage were analyzed. RESULT(S): Of 909 OCCCs collected from 92 cycles, 2.5%, 4.2%, 79.9%, and 13.4% were prophase I, metaphase I, metaphase II, and degenerating, respectively. No statistically significant differences were found in the percentage of intact metaphase II oocytes, the fertilization rate, or the cleavage rate among complexes with different morphologic grades. The morphologic grade of the OCCCs of transferred embryos in the pregnant group was not different from that in the nonpregnant group. CONCLUSION(S): Most oocytes were in metaphase II at the time of retrieval after ovarian stimulation. However, no relation was observed between the OCCC morphologic grade and oocyte nuclear maturity, the fertilization rate, or embryo cleavage. These observations suggest that OCCC morphology grading is a poor marker of oocyte quality.     

Cytogenetic analysis of human oocytes remaining unfertilized after intracytoplasmic sperm injection

Oocytes remaining unfertilized after intracytoplasmic sperm injection showed 12.0% aneuploidy (nondisjunction + unbalanced predivision), 3.4% structural aberrations, and 8.5% balanced predivision in fully karyotyped cells. However, the frequently observed complete or partial separation of chromatids, most probably caused by abortive activation, might complicate the evaluation of meiosis I-derived aneuploidy and questions the relevance of balanced predivision.     

Pregnancy after immature oocyte donation and intracytoplasmic sperm injection

Objective: To report a case of pregnancy from in vitro-matured primary oocytes fertilized by ICSI. The pregnancy occured in a woman who was in an oocyte donation program; the woman's husband had normal sperm parameters.

Design: Case report.

Setting: Private general hospital affiliated with a university hospital.

Patient(s): A recipient with premature ovarian failure, a recipient's husband with normal sperm, and a pregnant woman who donated her oocytes.

Intervention(s): Aspiration of immature oocytes during cesarean section, in vitro culture for maturation, ICSI of matured oocytes, coculture of fertilized oocytes.

Main Outcome Measure(s): Fertilization of oocytes by ICSI, and cleavage of embryos by Vero cell coculture.

Result(s): Two of seven immature oocytes became metaphase II oocytes, and both were fertilized by ICSI. The two zygotes were cocultured on Vero cells to become grade 1 two-cell embryos. Pregnancy was obtained after transfer.

Conclusion(s): More studies are necessary to clarify whether ICSI can increase the fertilization rate of in vitro-matured primary oocytes, and to clarify the role of coculture in fertilization.     

The use of piezo micromanipulation for intracytoplasmic sperm injection of human oocytes

Introduction: This paper describes the use of piezo actuator technology for intracytoplasmic sperm injection of human eggs. Methods: Piezo micromanipulation involves a simple and easily made injection pipette of very thin diameter and wall thickness, and it has recently been developed using a mouse model. No bevel or spike is required on the pipettes used. The piezo actuator attaches to conventional micromanipulators and acts to mechanically advance the pipette tip through the zona pellucida using a piezoelectric effect. The oolemma could, in some cases, also be broken using piezo. This technique was applied to 39 patients presenting for in vitro fertilization due to male-factor infertility. Results: The fertilization rate of eggs injected (60.5%), damage rate (7.6%), abnormality rate (3%), and ongoing pregnancy rate beyond 8 weeks (28.2%) are comparable to those for other techniques that have been successfully reported.     

Cytogenetic analysis of uncleaved oocytes after intracytoplasmic sperm injection

Purpose This work analyzes the causes of cleavage failure after intracytoplasmic sperm injection (ICSI) and the effect of the procedure on the chromosomes of the oocytes.Methods Ninety-seven uncleaved oocytes from 39 patients with severe male infertility or repeated IVF failure were fixed; 79 were analyzable. We checked the decondensation stage of spermatozoa nucleus and the chromosomal abnormalities of the oocytes.Results Among the fixed oocytes, the spermatozoa nucleus was present in 97% of the cases, and it was undecondensed in 89% of the cases, showing no evolution at all. A low rate (2.6%) of premature chromosome condensation (PCC) of the spermatozoa and a low rate (2.5%) of female diploïdy were observed. Among the oocytes that could be karyotyped, we observed a high rate (45%) of chromosome breakage.Conclusion ICSI fertilization failure was due mostly to the complete lack of evolution of the spermatozoa nucleus. Oocyte selection before ICSI seemed to lower the PCC rate. The high rate of oocyte chromosomal breakage rate has to be confirmed.     

Clinical and economic analysis of rescue intracytoplasmic sperm injection cycles

Objective.?To identify clinical and embryological factors that may predict success in rescue intracytoplasmic sperm injection (ICSI) cycles (after total fertilization failure has occurred) and to evaluate the cost effectiveness of rescue ICSI strategy. Additionally, follow-up of 20 rescue ICSI pregnancies is reported.

Design.?Retrospective analysis of total fertilization failure cycles.

Setting.?University-based tertiary medical center.

Material and methods.?In total, 92 patients who had undergone conventional in-vitro fertilization (IVF) cycles with total fertilization failure were included. The patients were divided into two subgroups: those who conceived through rescue ICSI and those who did not.

Results.?The pregnant members of the rescue ICSI subgroup were found to be significantly younger (32.9?±?4.2 vs. 36.3?±?4.5, respectively, p?=?0.0035,) and to have better-quality embryos than those who did not conceive (cumulative embryo score: 38.3?±?20.4 vs. 29.3?±?14.7, p?=?0.025). Cost effectiveness analysis showed 25% reduction in the cost per live birth when rescue ICSI is compared to cycle cancellation approach. The pregnancies follow-up did not show adverse perinatal outcome.

Conclusions.?Rescue ICSI is an option for salvaging IVF cycles complicated by total fertilization failure. Success in rescue ICSI was found to be associated with younger age and higher quality of embryos. Furthermore, the cost effectiveness of rescue ICSI in terms of total fertilization failure was found to be worthwhile.     

Cumulative ongoing pregnancy rate in intracytoplasmic sperm injection cycles

AIM: We investigated the cumulative probability of ongoing pregnancy in intracytoplasmic sperm injection (ICSI) cycles. METHODS: During a period of 6 years, 519 ICSI cycles were performed in 260 infertile couples at the Seoul National University Hospital. The cumulative rate of ongoing pregnancy (> or =12 weeks of gestation) was estimated by the Kaplan-Meier method. RESULTS: The cumulative ongoing pregnancy rate was 54.9% after six consecutive cycles of ICSI. The age of the female had a significant impact; after five consecutive cycles, cumulative ongoing pregnancy rate was 61.8% in < or =30 years of age, 51.7% in 31-35, and 15.3% in > or =36. There was no significant difference in the cumulative ongoing pregnancy rate between groups with severe male factor and previous unexplained low fertilization rate (< or =20%). Among the severe male factor group, cumulative ongoing pregnancy rate was not different according to the methods of sperm retrieval (surgically retrieved or ejaculated). CONCLUSION: Intracytoplasmic sperm injection could be applied successfully both to severe male factor and patients with previous unexplained low fertilization rate. The origin of spermatozoa does not affect ICSI outcome in terms of cumulative probability of ongoing pregnancy.     

Comparison of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection into human oocytes

OBJECTIVE: To compare the efficacy of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection (ICSI) into oocytes in patients with impaired semen parameters and no success with in vitro fertilization (IVF). STUDY DESIGN: A retrospective randomized study was conducted on 204 cycles for 104 couples with piezo-assisted ICSI and 122 cycles for 96 couples with conventional ICSI. Piezo-assisted ICSI consists of two steps, namely penetration of the zona pellucida alone with a piezo-pulse and then puncturing of the oolemma with a light negative pressure without piezo, as with conventional ICSI. The tips of injection pipettes were prepared after pulling by breakage with a scalpel under a microscope, so that the inner diameter at and near the tip was 5 microm, as for conventional ICSI. RESULTS: Piezo-assisted ICSI demonstrated significantly more favorable results, with a fertilization rate of 90.3% (conventional ICSI: 83.1%, p < 0.01) and a cleavage rate of 88.1% (conventional ICSI: 84.6%, p < 0.01). CONCLUSION: Piezo-assisted ICSI is easy to incorporate a spermatozoa exactly into the ooplasm with little deformation of the oocyte during insertion. Piezo-assisted ICSI can be used effectively for human oocytes to improve the fertilization, cleavage rates.     

Estradiol supplementation in intracytoplasmic sperm injection cycles with thin endometrium

Serial monitoring by plasma progesterone measurement is advised in the literature for fertility work-up ,to detect ovulation disturbances in women presenting with regular menstrual cycles. Three strategies to diagnose such ‘subtle ovulation disorders’ (SOD ,defined as anovulation ,inadequately timed ovulation or ovulation of a follicle of reduced size in regularly cycling women) were evaluated ,in order to investigate costs of such a diagnosis. On the basis of a ‘maximal’ ,an ‘ultrasound-only’ ,and a ‘preselection’ strategy ,total medical costs and costs including non-medical costs were calculated for each SOD diagnosis. A ‘maximal’ diagnostic strategy resulted in a total medical cost of ECU 9057 per diagnosis (including non-medical costs ECU 12 787); an ‘ultrasound-only’ strategy in ECU 4520 (ECU 6791) per diagnosis. By use of a ‘preselection’ strategy ,4.25% of the women were found to have an SOD ,at a cost of ECU 3036 (ECU 6868) for each diagnosis. As the real significance of SOD diagnosis for the prognosis of the patient to become pregnant without treatment remains unclear ,and as no randomized trials on treatment effectiveness have as yet been undertaken ,it is questionable whether this approach is worthwhile.     

不同来源精子行ICSI助孕1662个周期治疗结局分析

目的:探讨不同来源精子行卵胞浆内单精子显微注射(ICSI)助孕的妊娠结局。方法:回顾分析我中心2006年1月~2010年6月1662个ICSI治疗周期,按精子来源分为射出精子来源(重度少、弱精子)组1208周期,附睾穿刺取精(PESA)组324周期,睾丸穿刺取精(TESA)组130周期,比较3组胚胎发育情况和妊娠结局等指标。结果:射出精子组及PESA组受精率、卵裂率及2PN率较TESA组高(79.1%,77.9%vs 73.9%;98.7%,98.8%vs 96.6%;74.6%,73.0%vs 69.5%),TESA组1PN率较射出精子组及PESA组高(3.8%vs 2.2%,2.6%),差异均有统计学意义(P<0.05);3组优质胚胎率、胚胎种植率、临床妊娠率、异位妊娠率、流产率、单胎出生率、双胎出生率、畸形率无统计学差异。结论:PESA及TESA来源精子行ICSI助孕可获得与射出精子相似的妊娠结局。