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A new method for freezing testicular biopsy sperm: three pregnancies with sperm extracted from cryopreserved sections of seminiferous tubule.

OBJECTIVE: To determine whether spermatozoa, located in the seminiferous tubules obtained by needle puncture testicular biopsy, could be cryopreserved successfully within the tubules and subsequently be used for in oocyte fertilization via intracytoplasmic sperm injection (ICSI) after the spermatozoa were removed from the thawed tubules. DESIGN: Clinical case series. SETTING: Private IVF unit. PATIENT(S): Six azoospermic patients (four obstructive, two maturation arrest). MAIN OUTCOME MEASURE(S): Survival rate of thawed spermatozoa, fertilization rate of oocytes after ICSI with spermatozoa extracted from thawed tubules and pregnancies. RESULT(S): All six patients had adequate motile spermatozoa extracted from the thawed tubule sections, and all patients achieved fertilization via ICSI with their partner's eggs. The fertilization rate was 46%, compared with 56% obtained in other previous patient cycles using fresh testicular spermatozoa. Three pregnancies resulted from five ETs. CONCLUSION(S): Cryopreservation and subsequent thawing of seminiferous tubules proved to be a simple and successful method for storage of testicular spermatozoa, reducing the need for repetitive testicular biopsies and increasing the likelihood of sperm availability on the day of oocyte retrieval.

Adult↗

Microsurgical testis biopsy: a novel technique for retrieval of testicular tissue.

PURPOSE: In vitro fertilization with intracytoplasmic sperm injection has resulted in a dramatic increase in the need for diagnostic and therapeutic testis biopsies. We developed a microsurgical testis biopsy technique which allows identification of testicular vessels and individual seminiferous tubules. We compare the results of this technique to our prior series of nonmicroscopic biopsies. MATERIALS AND METHODS: A retrospective study of 226 consecutive patients who had undergone open testes biopsy with or without an operating microscope was performed. Between 1988 and 1994 standard open testis biopsy was performed without a microscope in 119 patients and a single sample of testicular tissue was taken. After 1994 microsurgical biopsy was performed under 6 to 25x magnification in 107 patients, nearly half of whom had multiple biopsies of each testis. The complication rates of the 2 procedures were compared. RESULTS: Scrotal hematoma required surgical drainage in 3 of the 119 standard testis biopsy cases and testis atrophy was noted in 1, for a total complication rate of 3.4%. There were no episodes of clinically detectable testicular atrophy or scrotal hematoma requiring surgical drainage in the 107 microsurgical biopsy cases (p<0.05). In 2 men the microscope allowed identification of larger tubules that contained sperm. CONCLUSIONS: Use of the operating microscope for testicular biopsy allows identification and avoidance of testicular vessels, minimizing complications. It also may allow selection of seminiferous tubules more likely to contain sperm.

Biopsy↗

Simplifying assisted conception techniques to make them universally available--a view from India.

In-vitro fertilization and related assisted conception techniques are beyond the reach of most infertile couples in India because they are so expensive. Recent innovations such as the use of the natural cycle, the techniques of gamete intra-Fallopian transfer, intravaginal culture and transcervical oocyte--sperm transfer, vaginal ultrasound-guided oocyte retrieval and the availability of ready-to-use culture medium have helped to simplify assisted conception. Research today should focus on further developing these techniques so that better pregnancy rates can be achieved with them. These simplified inexpensive assisted conception techniques can then be adapted for conditions in the developing world, so that infertile patients the world over can benefit from them.

Carbon Dioxide↗

Clinical review 87: In vitro fertilization for male factor infertility.

Since the first U.S. report of a successful delivery from in vitro fertilization in 1982 (65), progress in the field of assisted reproduction and micromanipulation has been truly dramatic. Perhaps the most exciting advances have been in the area of male factor infertility. Couples who previously would have been offered donor insemination or adoption are now achieving pregnancies despite severe impairments in semen quality, the presence of only single numbers of sperm in the ejaculate, or unreconstructable reproductive tract obstruction. Techniques of micromanipulation that were revolutionary less than five yr ago are now obsolete, replaced by even more successful methods. Even nonobstructive azoospermia resulting from maturation arrest or other impairments in germ cell development have been added to the list of treatable factors in male infertility, as sperm can frequently be extracted directly from testicular parenchyma that is aspirated or surgically biopsied. For patients without sperm in the testicular parenchyma, round spermatid or secondary spermatocyte injections are at least theoretically possible. Several important questions remain with regard to IVF-ICSI. 1) What should be the specific indications for IVF and IVF-ICSI? Should IVF alone ever be used for male factor infertility? 2) What are the reasons for failure to achieve pregnancy after ICSI, which still represent over half of our attempts at achieving ongoing pregnancies? 3) Can we be certain that using severely impaired or less mature sperm will not result in significant birth defects or in genetic abnormalities that could affect the offspring in adolescence or adulthood? 4) What is the most successful and cost effective approach for the infertile couple with impaired semen parameters? For couples with male factor infertility, careful evaluation and treatment of the man should be considered before assisted reproduction, including ICSI. Contemporary application of ICSI for severe male factor infertility can allow pregnancy rates up to 52% (33), with ongoing pregnancy and live delivery rates as high as 37% per IVF cycle attempt (27). As long as viable sperm are present in the ejaculate or retrievable from the reproductive tract, then ICSI procedures can be applied.

Congenital Abnormalities↗

Use of electroejaculation in the treatment of ejaculatory failure secondary to diabetes mellitus.

OBJECTIVES: To describe the experience of two male fertility programs using electroejaculation (EEJ) in the management of men with ejaculatory failure secondary to diabetes mellitus. METHODS: Twenty-nine EEJ procedures were performed in 7 diabetic men with ejaculatory failure. Results were reviewed with attention paid to sperm characteristics in both antegrade and retrograde specimens as well as pregnancy rates. RESULTS: Retrograde semen specimens retrieved from the bladder following EEJ contained a mean of 3444.5 million sperm (range 269.2 to 4996 million). Antegrade specimens contained a mean of 698.8 million sperm (range 226.8 to 1961 million). Mean sperm motility was 4% for retrograde specimens (range 0% to 11%) and 7% for antegrade specimens (1% to 15%). In all but 1 case, semen specimens were used for intrauterine insemination. The total number of motile sperm contained in the processed, inseminated specimens ranged from 1 to 87.2 million. In 1 case, the sperm obtained through EEJ was used in an in vitro fertilization procedure. CONCLUSIONS: EEJ can be successfully used to obtain sperm from men with ejaculatory failure due to diabetes mellitus. The procedure requires general anesthesia, and pregnancy rates after intrauterine insemination with the processed sperm are low. Advanced reproductive technologies may offer a feasible alternative, providing higher success rates with fewer procedures.

Diabetes Complications↗

Rapid decline in ovarian reserve after repeated ART cycles in a 22-year-old IVF patient.

OBJECTIVE: The objective was to discuss the rapid decline in ovarian reserve due to repeated assisted reproductive technologies (ART) in a 22-year-old patient. METHODS: This case report presents an infertile couple admitted to our IVF unit with the diagnosis of recurrent implantation failure and congenital uterine anomaly. Main outcome measures were the number of growing follicles and metaphase II oocytes retrieved. RESULTS: After recurrent implantation failure of three in vitro fertilization cycles, the couple underwent three consecutive intracytoplasmic sperm injection cycles in our center. With a high number of oocytes retrieved, first cycle resulted in implantation failure followed by an embryo transfer cancelled cycle. The third cycle was cancelled due to an unexpected poor response even with the highest gonadotrophin dose. CONCLUSION: Rapid decrease in ovarian reserve should be considered when offering repeated ART cycles even if the woman is very young and has a good ovarian reserve.

Adult↗

Assisted reproductive technology in the United States: 2000 results generated from the American Society for Reproductive Medicine/Society for Assisted Reproductive Technology Registry.

OBJECTIVE: To summarize the procedures and outcomes of ART initiated in the United States in 2000. DESIGN: Data were collected electronically using the SART Clinical Outcome Reporting System software and submitted to the American Society for Reproductive Medicine/ Society for Assisted Reproductive Technology Registry. PARTICIPANT(S): Three hundred eighty-three programs submitted data on procedures performed in 2000. Data were collated after November 2000 so that the outcome of all pregnancies established would be known. MAIN OUTCOME MEASURE(S): Incidence of clinical pregnancy, ectopic pregnancy, abortion, stillbirth, and delivery. RESULT(S): Programs reported initiating 99,989 cycles of ART treatment. Of these, 73,406 cycles involved fresh nondonor IVF (46.6% with intracytoplasmic sperm injection [ICSI]), with a delivery rate per retrieval of 29.9%; 549 were cycles of gamete intrafallopian transfer, with a delivery rate per retrieval of 24.7%; 763 were cycles of zygote intrafallopian transfer, with a delivery rate per retrieval of 29.9%. The following additional ART procedures were also initiated: 7,581 fresh donor oocyte cycles, with a delivery rate per transfer of 43.7%; 13,083 frozen embryo transfer procedures, with a delivery rate per transfer of 20.4%; 2,721 frozen embryo transfers using donated oocytes or embryos, with a delivery rate per transfer of 23.5%, and 1,200 cycles using a host uterus, with a delivery rate per transfer of 35.8%. In addition, 326 cycles were reported as combinations of more than one treatment type, 41 cycles as research, and 319 as embryo banking. As a result of all procedures, 25,394 deliveries were reported, resulting in 35,345 neonates, of which 35,031 were live born and 314 stillborn. CONCLUSION(S): In 2000, there were more programs reporting ART treatment and a significant (13.5%) increase in reported cycles compared to 1999. In comparable cycle types, overall success rate (deliveries per retrieval) exhibited an actual increase of 0.6%, which represents an increase of 2.2% when compared to the success rate for 1999.

Embryo Transfer↗

Pregnancies after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa.

In 25 patients (14 suffering from obstructive azoospermia, six from non-obstructive azoospermia, three from asthenoazoospermia and two from absence of ejaculation) spermatozoa were extracted from testicular biopsies. Intracytoplasmic sperm injection (ICSI) with fresh testicular spermatozoa was performed in 18 cases; spermatozoa in excess were cryopreserved in pills. No pregnancies were achieved. In the remaining seven patients, testicular spermatozoa were retrieved and cryopreserved during a diagnostic testicular biopsy. After thawing, sperm motility was assessed in 17 cases (68%), and 18 ICSI with cryopreserved testicular spermatozoa were performed. The mean two-pronuclear (2PN) fertilization rate was 59%, the mean cleavage rate was 92%, and six clinical pregnancies were achieved, all of them still ongoing (pregnancy rate 33%). A comparison of the results of ICSI carried out with fresh or cryopreserved testicular spermatozoa showed that the mean 2PN fertilization rates per cycle (53 compared with 55%), mean cleavage rates per cycle (99 compared with 96%) and embryo quality were not significantly different. In conclusion, cryopreservation of testicular spermatozoa is feasible, even in patients with non-obstructive azoospermia, and the results of ICSI with frozen-thawed testicular spermatozoa are similar to those obtained using fresh testicular spermatozoa. Cryopreservation of testicular spermatozoa may avoid repetition of testicular biopsies to retrieve spermatozoa for successive ICSI cycles in patients in whom the only source of motile spermatozoa is the testicle.

Adult↗

Comparative auto-controlled study between swim-up and Percoll preparation of fresh semen samples for in-vitro fertilization.

This study compared swim-up and Percoll preparation of fresh semen samples for in-vitro fertilization. Sixty trials of in-vitro fertilization (IVF), 38 with normal semen and 22 with abnormal semen, comprising 734 oocytes were included in the study. Each semen sample was prepared by both a swim-up technique and a simplified discontinuous (50%, 70%, 90%) Percoll gradient. The oocytes for each trial were distributed at random between the two sperm preparations and incubated with the same number of motile spermatozoa. Percoll gradient preparation produced a significantly higher final concentration of spermatozoa than swim-up preparation (mean +/- SEM: 6.6 +/- 1.5 x 10(6)/ml versus 1.9 +/- 0.2 x 10(6)/ml; P less than 0.01) but a significantly lower sperm motility (69 +/- 2% versus 94 +/- 1%; P less than 0.001) and a lower number of normal forms (55 +/- 2% versus 64 +/- 2%; P less than 0.01). The ability of the Percoll gradient method to extract motile spermatozoa was higher than that of the swim-up technique (20 +/- 15.6% versus 0.8 +/- 13.6%). Nevertheless, the rates of fertilization (61%), fertilization failure (18%) and polyspermia (9%), embryo quality evaluated by mean embryo scores (3.8 +/- 0.3) and the mean number of spare embryos frozen per trial (1.4 +/- 0.3) were strictly identical in both groups. The 24 pregnancies (including three from frozen--thawed embryos) obtained in these 60 trials (40% per oocyte retrieval) could not be separated according to the sperm preparation method, as embryos from both groups were replaced together.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Separation↗

Nuclear status and cytogenetics of embryos derived from in vitro-matured oocytes.

OBJECTIVE: To analyze embryos after in vitro maturation by investigating their nuclear status and cytogenetic constitution. DESIGN: Prospective randomized laboratory study. SETTING: Reproductive medicine unit in an academic hospital. PATIENT(S): Patients with male and tubal factor infertility undergoing fertility treatment. INTERVENTION(S): Denuded immature oocytes (n = 75) were matured in vitro for 24-30 hours, and intracytoplasmic sperm injection was performed 30 hours after oocyte retrieval. Fluorescence in situ hybridization was performed on the produced embryos. MAIN OUTCOME MEASURE(S): Blastomere content of the total embryo. RESULT(S): The in vitro-matured oocytes showed a similar fertilization rate as the in vivo-matured oocytes, but with a higher incidence of noncleavage (21.0%). In addition, 26.7% of these embryos arrested at the first mitotic division. Thirty embryos were processed for fluorescence in situ hybridization; only 6.7% had all mononuclear blastomeres, 30.0% had at least one binuclear blastomere, 43.3% had at least one multinuclear blastomere, and 56.6% contained anuclear cells. The chromosomal constitution was analyzed in 14 embryos, and chromosomal anomalies were found in 11 (78.5%). CONCLUSION(S): Germinal vesicle oocytes retrieved from superovulated patients and cultured in vitro for a short time had the ability to resume meiosis and achieve fertilization. However, arrest of embryo development was common. These embryos showed a high incidence of multinuclear blastomeres and aneuploidy, suggesting abnormal cytokinesis or genetic abnormalities.

Adult↗

Identification, proteomic profiling, and origin of ram epididymal fluid exosome-like vesicles.

Small membranous vesicles, between 25- and 75-nm diameter, were collected by high-speed centrifugation from the ram cauda epididymal fluid and were found to be normal constituents of this fluid and of the seminal plasma. The SDS-PAGE protein pattern of these vesicles was specific and very different from that of the caudal fluid, seminal plasma, sperm extract, and cytoplasmic droplets. After two-dimensional electrophoresis separation and mass spectrometry analysis, several proteins were identified and grouped into i) membrane-linked enzymes, such as dipeptidyl peptidase IV (DPP-IV), neprilysin (NEP), phosphodiesterase-I (E-NPP3), and protein G-beta; ii) vesicle-associated proteins, such as lactadherin (MFEG8-PAS6/7) and vacuolar ATPase; iii) several cytoskeleton-associated proteins, such as actin, ezrin and annexin; and iv) metabolic enzymes. The presence of some of these proteins as well as several different hydrophobic proteins secreted by the epididymis was further confirmed by immunoblotting. These markers showed that the majority of the vesicles originated from the cauda epididymal region. The physical and biochemical characteristics of these vesicles suggest they are the equivalent of the exosomes secreted by several cell types and epithelium. The main membrane-linked proteins of the vesicles were not retrieved in the extract from cauda or ejaculated sperm, suggesting that these vesicles did not fuse with sperm in vivo.

Amino Acid Sequence↗

Use of electively cryopreserved microsurgically aspirated epididymal sperm with IVF and intracytoplasmic sperm injection for obstructive azoospermia.

OBJECTIVE: To investigate the efficacy of using intentionally cryopreserved epididymal sperm in selected cases of obstructive azoospermia. DESIGN: A retrospective, nonrandomized study. SETTING: Academic research environment. PATIENTS: One hundred forty-one couples undergoing first-time IVF/ICSI using either fresh or cryopreserved epididymal sperm. INTERVENTIONS: The epididymides were microsurgically aspirated. MAIN OUTCOME MEASURES: Clinical pregnancy rates. RESULTS: Motile sperm were obtained from all men. For the fresh group, the mean total sperm aspirated was 99 x 10(6) with 5.5 vials frozen per patient after ICSI and 82 x 10(6) with 4.7 vials frozen per patient in the cryopreserved group. No statistically significant difference in oocyte fertilization rate or number of embryos transferred was noted between groups. Of 108 patients using freshly aspirated sperm, 72 (66.7%) achieved clinical pregnancy. Of 33 patients in the group using cryopreserved sperm, 20 (60.6%) achieved clinical pregnancy (P=0.47). CONCLUSIONS: In selected ideal cases of unreconstructable azoospermia, elective open microsurgical epididymal sperm aspiration with cryopreservation yields pregnancy rates similar to that employing fresh sperm. The advantages of this method are: (1) Use of cryopreserved sperm obviates the logistics problems associated with the use of fresh sperm. (2) Abundant high-quality sperm can be cryopreserved in a single procedure for all future attempts at IVF/ICSI. Rarely, viable sperm will not be present after thawing, and fresh retrieval will be necessary.

Cryopreservation↗

Outcome of intracytoplasmic sperm injection in azoospermic patients: stressing the liaison between the urologist and reproductive medicine specialist.

OBJECTIVES: To analyze the outcome of intracytoplasmic sperm injection (ICSI) cycles in infertile couples in whom the main diagnosis of infertility was azoospermia of obstructive and nonobstructive origin. METHODS: Eighty-three consecutive ICSI cycles were carried out with retrieved testicular or epididymal spermatozoa, 60 cycles in 32 patients with obstructive azoospermia and 23 cycles in 12 patients with nonobstructive azoospermia. Fifty-four testicular biopsies (testicular sperm extraction) and 18 epididymal aspirations (microepididymal sperm aspiration) were performed.Results. Motile spermatozoa were recovered in 65 cycles (90.3%). In another 3 (4.2%), nonmotile spermatozoa were retrieved. In 4 patients (5.5%), sperm could not be recovered. In 11 cycles, frozen sperm from a previous procedure were used. A significantly lower fertilization rate (64% versus 73%, P = 0.02), clinical pregnancy rate (13% versus 47%, P <0.001), and good embryo quality rates (35% versus 56%, P = 0.009) were observed in patients with nonobstructive azoospermia. In patients with obstructive azoospermia, no significant differences were observed when the outcome was analyzed on the basis of the sperm origin (ie, from testicular sperm extraction or microepididymal sperm aspiration). CONCLUSIONS: When combining testicular sperm extraction or microepididymal sperm aspiration with ICSI in patients with obstructive azoospermia, the results in terms of fertilization, implantation, and pregnancy rates were similar to those found in patients with nonazoospermic obstruction who underwent ICSI with ejaculated sperm. Patients with nonobstructive azoospermia had lower fertilization, embryo quality, and pregnancy rates than did those with obstructive azoospermia, probably because of severe defects in spermatogenesis, leading to poor gamete quality. The urologist and reproductive endocrinologist now have an excellent therapeutic option to offer men with previously intractable infertility.

Adult↗

[Impact of high response of ovary to gonadotropin stimulation on the outcome of in vitro fertilization and embryo transfer or intracytoplasmic sperm injection].

OBJECTIVE: To evaluate the impact of elevated peak estradiol (E(2)) levels and a high number of retrieved oocytes on implantation and pregnancy rate in patients undergoing in vitro fertilization-embryo transfer (IVF-ET) and intracytoplasmic sperm injection (ICSI). METHODS: Retrospectively analyzed 474 infertile women undergoing 510 cycles for IVF-ET/ICSI treatment during the period of March 1999 to December 2000. Using a standard long protocol/flare-up protocol [(gonadotropic hormone releasing hormone agonist/high pure follicle-stimulating hormone (FSH-HP)/human chorionic gonadotropin (hCG)] for ovarian stimulation. High responders were defined as those who had peak E(2) levels of > 11 010 pmol/L on the day of hCG administration (n = 160) or > 15 retrieved oocytes (n = 148). Normal responders were defined as those who had peak E(2) levels of </= 11 010 pmol/L on the day of hCG administration (n = 350) or </= 15 retrieved oocytes (n = 362). RESULTS: There were statistically significant differences in age, infertile years, basal FSH level and the ampules of FSH-HP required between high and normal responders (P < 0.01 and P < 0.05). Ovarian hyperstimulation syndrome (OHSS) increased significantly in the high responders (P < 0.01). There were no statistically significant differences in implantation rate or pregnancy rate between high and normal responders. CONCLUSION: Elevated peak E(2) levels and high oocytes retrieved were not detrimental to IVF outcome, but had high incidence of OHSS.

Adult↗

Seven pregnancies and deliveries from non-mosaic Klinefelter syndrome patients using fresh and frozen testicular sperm.

PURPOSE: The aim of this study was to investigate the feasibility of using frozen-thawed testicular sperm as well as the timing of testicular sperm extraction (TESE) in patients with non-mosaic Klinefelter syndrome. METHODS: Intracytoplasmic sperm injection (ICSI) was performed in six of 17 (35%) patients whose sperm was recovered by TESE. Multiple biopsies of both testes were performed on the day of oocyte retrieval in all but one of the six patients. RESULTS: Seven pregnancies and deliveries were achieved in five couples, and one couple was unsuccessful. Five pregnancies were achieved using fresh motile sperm, and two were achieved using frozen-thawed sperm. Sperm cryopreservation was not possible in one of the five couples because of the small number of recovered sperm, and possible in four other couples for subsequent ICSI. One woman whose husband had TESE performed prior to ovarian stimulation did not become pregnant. This may be due to the attainment of only a few immotile sperm following the frozen-thawed procedure. CONCLUSION: The outcome of ICSI using fresh or frozen-thawed testicular sperm in patients with non-mosaic Klinefelter syndrome was identical; however, TESE should be performed on the day of oocyte retrieval until such time as a procedure with a higher sperm yield from TESE is available. Moreover, an improved recovery procedure after cryopreservation-thawing of a single spermatozoon must be developed.

Adult↗

Analysis of intracytoplasmic sperm injection procedures related to delayed insemination and ejaculated, epididymal and testicular spermatozoa.

In all, 1210 treatment cycles were divided into three categories for retrospective analysis according to the period of delay between oocyte retrieval (occurring at a fixed time after human chorionic gonadotrophin) and intracytoplasmic sperm injection (ICSI) of <3 h, 3-5 h, >5 h (referred to as 'delayed ICSI'). Three stages from oocyte to the birth of a live baby were identified for statistical analysis, (i) fertilization (2PN zygotes), (ii) cleavage of 2PN zygotes, (iii) transferred embryo to live birth. Stages 1, 2 and 3 were analysed statistically for the three time periods. Chi-square analysis showed no significant effect of delayed ICSI on fertilization (chi(2) = 3.615, P = 0.65), and embryo transfer to birth (chi(2) = 1.840, P = 0.399). The effect on cleavage was significant (chi(2) = 9.625, P = 0.008). However, shorter incubation times produced results which were better than the traditional longer ones. The success rate at the cleavage stage was so high that the marginal advantage had very little effect on the overall process. This study of a substantial patient sample establishes that ICSI on a peri-ovulatory oocyte (<3 h after oocyte retrieval) does not compromise outcome parameters, and that longer periods of incubation (>5 h) do not offer a statistically significant advantage.

Journal Article↗

Day three versus day two embryo transfer following in vitro fertilization or intracytoplasmic sperm injection.

BACKGROUND: Embryo transfer (ET) was traditionally performed two days after oocyte retrieval; however, developments in culture media have allowed embryos to be maintained in culture for longer periods. Delaying transfer from day two to day three would allow for further development of the embryo and might have a positive effect on pregnancy outcomes. OBJECTIVES: To determine if there is any difference in live birth and pregnancy rates when ET is performed on day three, compared with day two, in infertile couples undergoing treatment with in vitro fertilization (IVF), including intracytoplasmic sperm injection (ICSI). SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders & Subfertility Group trials register (17th December 2003), the Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 1, 2003), MEDLINE (1966 to 2003), EMBASE (1980 to 2003) and Biological Abstracts Databases (1980 to 2003), the National Research Register (NRR), the Medical Research Council's Clinical Trials Register, the NHS Centre for Reviews and Dissemination, citation lists of relevant publications, review articles and included studies, as well as abstracts of appropriate scientific meetings. SELECTION CRITERIA: Randomized controlled trials that compared day three versus day two embryo transfer after oocyte retrieval during an IVF or ICSI treatment cycle in subfertile couples. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Study authors were contacted for additional information. The primary outcome measures were live birth rate and ongoing pregnancy rate. MAIN RESULTS: Ten studies involving 2027 women were included, but only three studies reported live birth and four reported ongoing pregnancy rates. The pooled odds ratios (day three compared to day two) were 1.07, 95% CI 0.84 to 1.37 for live birth and 1.05, 95% CI 0.83 to 1.32 for ongoing pregnancy. From ten studies, the pooled odds ratio for clinical pregnancy was 1.26, 95% CI 1.06 to 1.51. Sub-group analysis revealed that this advantage occurred in those undergoing ICSI cycles. A higher miscarriage rate with day 3 ET in ICSI cycles works to negate the increased clinical pregnancy rate, in agreement with the finding of no significant difference between treatments in live birth rate. REVIEWERS' CONCLUSIONS: Although an increase in clinical pregnancy rate with day three embryo transfer was demonstrated, at present there is not sufficient good quality evidence to suggest an improvement in live birth when embryo transfer is delayed from day two to day three.

Embryo Transfer↗

Monochorionic-triamniotic triplet pregnancy after intracytoplasmic sperm injection, assisted hatching, and two-embryo transfer: first reported case following IVF.

BACKGROUND: We present a case of monochorionic-triamniotic pregnancy that developed after embryo transfer following in vitro fertilization (IVF). METHODS: After controlled ovarian hyperstimulation and transvaginal retrieval of 22 metaphase II oocytes, fertilization was accomplished with intracytoplasmic sperm injection (ICSI). Assisted embryo hatching was performed, and two embryos were transferred in utero. One non-transferred blastocyst was cryopreserved. RESULTS: Fourteen days post-transfer, serum hCG level was 423 mIU/ml and subsequent transvaginal ultrasound revealed a single intrauterine gestational sac with three separate amnion compartments. Three distinct foci of cardiac motion were detected and the diagnosis was revised to monochorionic-triamniotic triplet pregnancy. Antenatal management included cerclage placement at 19 weeks gestation and hospital admission at 28 weeks gestation due to mild preeclampsia. Three viable female infants were delivered via cesarean at 30 5/7 weeks gestation. CONCLUSIONS: The incidence of triplet delivery in humans is approximately 1:6400, and such pregnancies are classified as high-risk for reasons described in this report. We also outline an obstetric management strategy designed to optimize outcomes. The roles of IVF, ICSI, assisted embryo hatching and associated laboratory culture conditions on the subsequent development of monozygotic/monochorionic pregnancy remain controversial. As demonstrated here, even when two-embryo transfer is employed after IVF the statistical probability of monozygotic multiple gestation cannot be reduced to zero. We encourage discussion of this possibility during informed consent for the advanced reproductive technologies.

Journal Article↗