Chromosome analysis of blastocysts derived from single pronuclear zygotes by array CGH and clinical outcomes by the transfer of single pronuclear zygotes.

PURPOSE: This study aimed to explore the aneuploidy of blastocysts derived from single pronuclear (1PN) zygotes, almost 75% of which were regarded as diploid, using array CGH and examine the pregnancy outcomes. METHODS: Embryonic aneuploidy screening of sixteen embryos from 1PN zygotes and sixteen embryos from 2PN zygotes was performed using array CGH in study 1. In addition, the reproductive outcome of 1761 single blastocysts, after untested frozen-thawed blastocyst transfer in IVF/ICSI patients, was retrospectively analyzed and compared between the 1PN and 2PN groups in study 2. RESULTS: The aneuploidy rates were 30.8% (4/13) in 1PN IVF, 33.3% (1/3) in 1PN ICSI, 46.2% (6/13) in 2PN IVF, and 100% (3/3) in 2PN ICSI. The 1PN group achieved clinical pregnancy in 25.0% (7/28) of IVF and 30.0% (3/10) of ICSI, whereas these rates in the 2PN control group were 44.6% (557/1250) of IVF and 37.4% (177/473) of ICSI. No miscarriage occurred in the pregnancies from 1PN zygotes, whereas the rates of miscarriage in the 2PN control group were 22.6% (126/557) in IVF and 22.2% (39/176) in ICSI. The delivery rate was similar in all groups. Ten deliveries in the 1PN group showed no newborn malformation. CONCLUSION: Within the limits of the small sample size, our results suggest that the aneuploidy and delivery rates of the blastocysts derived from 1PN zygotes are the same as those derived from 2PN zygotes. Blastocysts derived from 1PN zygotes may be used clinically and could increase the chance of pregnancy.

Prediction of pregnancy rate by blastocyst morphological score and age, based on 1,488 single frozen-thawed blastocyst transfer cycles.

OBJECTIVE: To present an estimation of the pregnancy rate after IVF or intracytoplasmic sperm injection stratified by blastocyst morphology and age. DESIGN: Retrospective analysis. SETTING: Private IVF clinic. PATIENT(S): A total of 1,488 single frozen-thawed blastocyst transfer cycles. INTERVENTION(S): All frozen-thawed blastocysts used in the study were obtained in the patients' first oocytes retrieval cycles. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate (CPR), viable pregnancy rate (VPR), and delivery rate (DR) according to blastocyst morphological score (Gardner and Schoolcraft score) in three different age groups: women aged 22-33 years, 34-37 years, and 38-45 years. RESULT(S): The quality of blastocysts degraded as age group increased. The proportions of good-quality blastocysts (grades 4 and above) were 62.3%, 56.3%, and 41.1% in age groups of 22-33 years, 34-37 years, and 38-45 years. Within the same blastocyst quality, CPR, VPR, and DR tended to be lower with increased age. Chances of pregnancy were reduced by 60% or more for women 38 years and older with blastocyst morphology of grades 1 and 2. Significant trends were observed for both age and blastocyst morphology groups. CONCLUSION(S): There was a significant correlation between blastocyst quality and CPR, VPR, and DR in addition to the influence of age on the three rates. The findings may help predict successful pregnancy in single-blastocyst transfer.

The effects of severity of oligozoospermia on Intracytoplasmic Sperm Injection (ICSI) cycle outcome.

The objective of this study was to explore the relationship between the severity of oligozoospermia and the development of embryos and clinical outcome in patients undergoing ICSI. A total of 908 intracytoplasmic sperm injection cycles involving women of < or =37 years of age were included in this study. The patients were divided into four treatment groups according to the results of an analysis of their husbands' semen: (A) mild oligozoospermia that ranged from 10 x 10(6)/ml to <20 x 10(6)/ml (n=283), (B) mild to severe oligozoospermia that ranged from 5 x 10(6)/ml to <10 x 10(6)/ml (n=192), (C) severe oligospermia that ranged from 1 x10(6)/ml to <5 x 10(6)/ml (n=259), and (D) very severe oligozoospermia that ranged from 0 to <1 x10(6)/ml (n=174). Two pronuclei (PN) oocytes at MII were injected and the development of high quality embryos on day 2, as well blastocyst formation rate on day 5, the implantation rate, clinical pregnancies, and fetal loss, were examined. A lower percentage of two pronuclei (2PN) oocytes in the very severe oligozoospermia group was observed, however, there was no difference in clinical outcome when the oligozoospermic patients were divided by sperm concentration. In addition, no significant difference was detected in zygote production or clinical outcome between spermatozoa with a motility of <40% and spermatozoa with a motility of > or =40%. The results of this study emphasize the importance of selecting good quality sperm for oocyte injection, especially in cases involving very severe oligozoospermia.

A healthy birth after intracytoplasmic sperm injection using ejaculated spermatozoa from a patient with Kartagener's syndrome.

OBJECTIVE: To report a healthy birth that was achieved by intracytoplasmic sperm injection (ICSI) with use of ejaculated spermatozoa from a patient with Kartagener's syndrome. DESIGN: Case report. SETTING: Private infertility clinic. PATIENT(S): Couple with male factor infertility due to Kartagener's syndrome. INTERVENTION(S): Intracytoplasmic sperm injection with ejaculated sperm. MAIN OUTCOME MEASURE(S): Semen characteristics, sperm motility, fertilization, pregnancy, and birth after ICSI. RESULT(S): With ejaculated sperm, the fertilization rates were 73% in the first stimulation cycle and 100% in the second cycle. Intracytoplasmic sperm injection was successful. The pregnancy resulted in birth of a single healthy child. CONCLUSION(S): With ejaculated sperm, successful pregnancy after ICSI in couples with Kartagener's syndrome is possible. Kartagener's syndrome is a heterogeneous group of disorders with similar clinical presentations, and treatment should be individualized depending on sperm motility.

Stimulation of endometrium embryo transfer can improve implantation and pregnancy rates for patients undergoing assisted reproductive technology for the first time with a high-grade blastocyst.

OBJECTIVE: To evaluate whether stimulation of endometrium embryo transfer (SEET) can improve implantation rate and pregnancy rate (PR) for patients undergoing assisted reproductive technology (ART) for the first time by injecting embryo culture supernatant into the uterus before blastocyst transfer (BT). DESIGN: Randomized, controlled trial. SETTING: Private in vitro fertilization clinic. PATIENT(S): Forty-eight women in the BT group, 48 women in the stimulation group who had culture medium injected into the uterus before BT, and 48 women in the SEET group. INTERVENTION(S): Injection of embryo culture supernatant and injection of culture medium. MAIN OUTCOME MEASURE(S): Implantation rates and PRs. RESULT(S): Odds ratios of successful implantation rate for stimulation and SEET in patients with high-grade blastocysts, having BT as reference, were 2.58 and 6.46 without adjustment, and 5.91 and 9.20 after adjusting for basal FSH levels and period of infertility. Odds ratios of clinical pregnancies were 2.47 and 4.32 without adjustment, and 4.46 and 5.10 with adjustment, respectively. In groups with low-grade blastocysts, such tendencies were not observed. CONCLUSION(S): The SEET may be an effective method for increasing implantation rate and PR for first-time ART patients who have a high-grade blastocyst.

Fertilization and pregnancy using cryopreserved testicular sperm for intracytoplasmic sperm injection with azoospermia.

OBJECTIVES: To report fertility for men with obstructive (OA) and nonobstructive (NOA) azoospermia after frozen and thawed spermatozoa recovered from the seminiferous tubules and intracytoplasmic sperm injection (ICSI) and to evaluate the factors other than spermatozoa. DESIGN: Retrospective clinical analysis. SETTING: Male infertility clinic for testicular sperm extraction and freezing/thawing (cryoTESE) and assisted reproductive technologies clinic for ICSI. PATIENT(S): Seventy-four men with OA and 140 men with NOA undergoing attempted cryoTESE-ICSI. INTERVENTION(S): Seventy-three couples with OA underwent a total of 184 cryoTESE-ICSI cycles, and 46 couples with NOA underwent a total of 75 cryoTESE-ICSI cycles. MAIN OUTCOME MEASURE(S): The numbers of eggs at metaphase II injected, two-pronuclei oocytes, normal cleaved embryos, embryos transferred, transfer cycles, biochemical pregnancies, and clinical pregnancies, as well as the implantation and delivery rates, were examined. RESULT(S): Fertilization rate in NOA was significantly lower than in OA. Neither the pathology, the source, nor the quantity of spermatozoa had any effect on fertilization or pregnancy rates. Maternal age had no effect on fertilization or embryo cleavage, but did dramatically affect the implantation, pregnancy, and delivery rates in NOA and OA. CONCLUSION(S): Good pregnancy rates were achieved without significant differences among the sperm sources. The pregnancy and the delivery rate were dependent strictly on the age of the female partner but not on her ovarian reserve.

Stimulation of endometrium embryo transfer (SEET): injection of embryo culture supernatant into the uterine cavity before blastocyst transfer can improve implantation and pregnancy rates.

OBJECTIVE: To evaluate whether stimulation of endometrium embryo transfer (SEET) by injecting embryo culture supernatant (ECS) into the uterus before blastocyst transfer (BT) can improve implantation and pregnancy rates. DESIGN: Randomized, controlled trial. SETTING: Private in vitro fertilization clinic. PATIENT(S): 23 women in the SEET group, and 25 women in the control group. INTERVENTION(S): Injection of ECS. MAIN OUTCOME MEASURE(S): Implantation rates, pregnancy rates, and serum beta human chorionic gonadotropin (beta-hCG) levels at 4 weeks and 1 day of gestation. RESULT(S): Communication between the embryo and the endometrium (cross-talk) plays an important role in implantation. Several embryonic factors that modulate endometrial receptivity have been detected in ECS. In BT, cross-talk is absent until the blastocysts are transferred, so we injected ECS into the uterine cavity before BT to stimulate the endometrium and provide an optimum environment for implantation of forthcoming blastocysts. The implantation rates, pregnancy rates, and beta-hCG levels in the SEET group were statistically significantly higher than those in the BT control group. CONCLUSION(S): A novel method of embryo transfer based on the concept of embryo-dependent induction of endometrial receptivity, SEET may be an effective option for increasing implantation and pregnancy rates.

Effectiveness of two-step (consecutive) embryo transfer in patients who have two embryos on day 2: comparison with cleavage-stage embryo transfer.

OBJECTIVE: To evaluate the effectiveness, for patients who have only two embryos on day 2, of a two-step (consecutive) embryo transfer (ET) procedure in which a cleaved embryo is transferred on day 2 and a single blastocyst is transferred on day 5. DESIGN: Observational comparative study. SETTING: Private IVF clinic. PATIENT(S): Ninety two-step ET cycles were performed in patients who had two embryos on day 2 (two-step group). Ninety day-2 ET cycles were performed in age- and infertility-matched patients who had two embryos on day 2 (control group). INTERVENTION(S): Cleaved-ET, extended culture of one embryo, and a second transfer of a blastocyst. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): The pregnancy and implantation rates in the two-step group (respectively 33.3% and 17.2%) were significantly higher than those in the control group (18.9% and 9.4%). Thirty-nine of the patients in the two-step group (43.3%) could not proceed to the second step of ET because no viable blastocyst could be obtained, but four of them conceived anyway. CONCLUSION(S): Taking advantage of both day-2 ET and blastocyst transfer, two-step ET may be an effective option for ET in patients who have an insufficient number of embryos.

Effectiveness of 2-step (consecutive) embryo transfer. Comparison with cleavage-stage transfer.

OBJECTIVE: To evaluate the effectiveness of 2-step (consecutive) embryo transfer (ET), in which two cleaved embryos are transferred on day 2 (first step of ET) and a single blastocyst is transferred on day 5 (second step of ET) in comparison with conventional day 2 ET. STUDY DESIGN: Observational comparative study at different time periods. Subjects were those who had > or = 4 embryos on day 2 after oocyte retrieval. Those in the conventional group (n = 60) received conventional day 2 ET, in which 3 cleaved embryos were transferred on day 2 between January 1999 and June 2000. Those in the 2-step group (n = 124) received 2-step embryo transfer, in which 2 cleaved embryos were transferred on day 2 and a blastocyst was transferred on day 5 between September 2000 and February 2002. Patients who planned to receive 2-step ET but could not proceed to the second step of ET because of failure to produce blastocysts were included in the 2-step group. RESULTS: The ongoing pregnancy rate with 2-step ET (51.6%) was significantly higher than with conventional ET (26.7%) (P < .01). The clinical pregnancy rate and implantation rate in conventional and 2-step ET were 30.0% vs. 59.7% and 11.1% vs. 27.8%, respectively (P < .001). CONCLUSION: Two-step ET may benefit patients who can obtain a sufficient number of embryos.