Average Number Of Embryos Research Articles

179 COMPARISON OF OOCYTE AND EMBRYO PRODUCTION AMONG BOS TAURUS, BOS INDICUS, AND INDICUS-TAURUS DONOR COWS

In recent years, Brazil has become the leading country in the world for the number of embryos produced in vitro (Thibier M 2009 IETS Embryo Transfer Newsletter 22, 12-19). This is partly due to the large numbers of Bos indicus animals in Brazil, making up about 80% of the total cattle. The mean oocyte production per ultrasound-guided follicular aspiration from Bos indicus is higher than those for European breeds (Pontes JHF et al. 2009 Theriogenology 71, 690-697). In the present study, we analyzed 5407 ovum pick ups (OPU) and compared the average production of total (n = 90,086) and viable (n = 64,826) oocytes and the number of embryos produced in vitro from Gir (Bos taurus indicus), Holstein (Bos taurus taurus), 1/4 Holstein × 3/4 Gir, and 1/2 Holstein-Gir crossbreed cows. To obtain oocytes, OPU was repeated from 4 to 7 times (mean = 5.7 ± 2.4) in each donor cow aged from 3 to 7 years (mean = 5.0 ± 2.3) during a 12-mo period. COCs (n = 90,086) obtained were classified according to the presence of cumulus cells and the oocyte cytoplasm aspect (homogeneous or heterogeneous/fragmented). The viable oocytes (n = 64,826) were in vitro matured for 24 h at 38.8°C in an atmosphere of 5% CO2 in air. Since this was a commercial programm, frozen sexed semen (2 × 106 mL-1) from Gir (n = 8) or Holstein (n = 7) sires previously tested for high efficiency was used for IVF. Fertilization was carried out (18-20 h) and the presumed embryos were cultured for 7 days in the same conditions as were used for IVM. Data were analyzed by ANOVA. On average, 16.7 ± 6.2 oocytes were obtained per OPU/IVF procedure and 71.96% were considered viable. The mean numbers of total oocytes per OPU/IVF procedure were 17.1 ± 4.4 for Gir cows (n = 617), 11.4 ± 3.9 for Holstein cows (n = 180), 20.4 ± 5.8 for 1/4 Holstein × 3/4 Gir (n = 44), and 31.4 ± 5.6 for 1/2 Holstein-Gir crossbreed females (n = 37, P < 0.01). The mean numbers of viable oocytes per OPU/IVF procedure were 12.1 ± 3.8 for Gir cows, 8.0 ± 2.6 for Holstein cows, 16.8, ± 5.0 for 1/4 Holstein × 3/4 Gir, and 24.3 ± 4.7 for 1/2 Holstein-Gir crossbreed females (P < 0.01). The average number of embryos produced by OPU/IVF were 3.2 (n = 12,243/3378) for Gir cows, 2.2 (n = 2426/1138) for Holstein cows, 3.9 (n = 1033/267) for 1/4 Holstein × 3/4 Gir, and 5.5 (n = 1222/224) for 1/2 Holstein-Gir. The average number of embryos produced per IVF session from 1/2 taurus × indicus donor cows was greater (P < 0.01) than from Bos indicus cows. The number of recoverable and viable oocytes and the number of embryos produced in vitro from Bos indicus donors were higher than from Bos taurus females. Therefore, the highest oocyte yield and the greatest embryo production were obtained from 1/2 taurus × indicus females. This work was supported by In Vitro Brasil.

SURVEY OF ROOF RAT Rattus rattus, (Linn.) IN EL-IBRAHEMIA DISTRICT, SHARKIA GOVERNORATE

Survey of the roof rat Rattus rattus (Linn.) by live traps in the three different locations, (houses, granaries and poultry farm) in El-Ibrahemia district, Sharkia Governorate were studied, during two successive years (2004/2005 and 2005/2006). The total number of R. rattus (Linn.) were (435 and 329 individuals), (84 and 67 individuals) and (86 and 66 individuals) recorded in houses, granaries and poultry farm during the 1st and 2nd years, respectively. The data indicated that females outnumbered males all over the year in the three studied locations except during the 2nd year, in the granaries. The number of mature stages exceeded that of the immature ones at the different locations during all the studied years. On the other hand, the poultry farm obtained the highest average number of embryos per pregnant female for the roof rat was 7.1 and 7.3 during the 1st and 2nd years, respectively.

Studies on genetic transformation of Theobroma cacao L.: evaluation of different polyamines and antibiotics on somatic embryogenesis and the efficiency of uidA gene transfer by Agrobacterium tumefaciens

In order to develop a more efficient genetic transformation system for cacao somatic embryos, the effects of polyamines and β-lactam antibiotics on somatic embryogenesis, hygromycin as selective agent, and different factors affecting uidA gene transfer have been evaluated. The polyamines putrescine, spermidine, and spermine significantly improved secondary somatic embryogenesis in cacao. Spermine at 1,000 μM provided the best responses, increasing 6.7× the percentage of embryogenic callus and 2.5× the average number of embryos per embryogenic callus. The β-lactam antibiotics timentin and meropenem, used for Agrobacterium tumefaciens counter-selection, had a non-detrimental effect on secondary somatic embryogenesis, depending on their concentration, whereas the commonly used β-lactam cefotaxime inhibited it, irrespective of the tested concentration. Hygromycin showed a strong inhibitory effect on secondary somatic embryogenesis of cacao, impairing completely the embryo production at 20 mg l−1. Following the criterion of GUS activity, the best conditions for T-DNA transfer into cotyledon explants from primary somatic embryos of cacao were a sonication of the explants for 100 s, a 20-min incubation period in Agrobacterium solution, an Agrobacterium concentration of 1.0 (OD600), and cocultivation of the explants on tobacco feeder layers. These findings will have important implications for studies on functional genomics of cacao.

ICMART report on global trends in ART from 2000-2004: how does the USA compare?

OBJECTIVE: To identify trends affecting efficacy and safety of ART through analysis of the International Committee Monitoring ART's (ICMART's) global database. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Global trends relating to efficacy and safety for 2000, 2002, 2003 and 2004 were identified. Similarities and differences among North America (NA), Europe (EU), Australia/New Zealand (ANZ), Latin America (LA), Middle East (ME), Asia (AS) and selected countries were compared with the USA. RESULTS: USA delivery rates per retrieval were stable from 2000 to 2004 at 31% while the EU improved from 16.4% to 22.5%. Cycles with 1 or 2 embryo transfer increased in USA from 34.5% to 49.4% and in EU from 61.5% to 74.6%. The average number of embryos transferred decreased in USA from 3.0 to 2.7. A global reduction of 3 to 2 embryos transferred in NA, EU and ANZ and from 4 to 3 embryos in LA, ME and AS occurred. The USA performed relatively fewer FET cycles but had higher pregnancy rates per transfer at 31.8%. For egg donation the USA doubled the number of cycles between 2000 and 2004 while the EU was stable, and increased delivery rates to over 50% which was double the EU. The USA reduced its triplet delivery rate from 4.3% to 2.6%, Canada 2.8% to 1.6%, the United Kingdom 2.3% to 0.3%, and Sweden 0.4% to 0%. The USA twin delivery rate dropped slightly from 31.1% to 30.4%. Only Sweden, with a major shift to single embryo transfer (SET), reduced the twin rate significantly, from 21.8% to 5.6%. CONCLUSIONS: From 2000 to 2004 delivery rates were maintained despite the reduced average number of embryos transferred. FET is performed more frequently as embryo transfer number is reduced. Its efficacy is important to maintain cumulative delivery rates. Triplet delivery rate is reduced by transfer of 2 rather than 3 embryos, but reduction of twin rate requires SET. International data provide important insight into ART efficacy and safety.

Unconfounding the effects of climate and density dependence using 60 years of data on spiny dogfish (Squalus acanthias)

The confounded effects of changes in climate and density on a population’s demography are hard to separate for long-lived species because demographic traits are usually the aggregated result of conditions faced over years. Demographic parameters are compared for spiny dogfish ( Squalus acanthias ) in the Northeast Pacific, the longest lived and latest maturing of all sharks, using samples from the 1940s and 2000s. This 60-year interval has seen ocean temperatures rise by almost 1 °C and dogfish harvests peak above 50 000 tonnes (t)·annum–1 and then drop below 50 t·annum–1. Over this period, the age at 50% maturity for dogfish declined from 43 to 32 years, while the estimated average number of embryos per litter for a 100 cm dogfish increased from 5.9 to 6.7. Growth parameters changed significantly, with faster growth to a smaller size. Leslie matrix analysis showed that these changes could lead to an increase in population growth rate of about 1%. Comparison with published demographic parameters from the 1970s and 1980s indicated that the greatest change in demographic parameters occurred between the 1940s and 1970s, prior to the largest changes in ocean temperatures. The implications for fishing on long-lived populations during times of rapid environmental change are explored.

Effect of sucrose concentration, polyethylene glycol and activated charcoal on maturation and regeneration of Abies cephalonica somatic embryos

For promoting the maturation of two embryogenic cell lines of Abies cephalonica Loud., the effect of two sucrose concentrations (87.6 and 175.2 mM) applied alone (Control) or in combination with activated charcoal (AC) (1 week) or with polyethylene glycol (PEG1 and PEG4) (1 or 4 weeks) was studied. The effect of each maturation medium was tested with four concentrations of abscisic acid (4, 8, 16 and 32 μM ABA). AC supplement significantly increased the percentage of embryogenic cell masses (ECMs) with cotyledonary embryos but did not affect the average number of embryos per 1 g of ECMs fresh weight. The highest percentages of ECMs (77.5%) with cotyledonary somatic embryos and the highest average number of somatic embryos (36.5 ± 13.4) were found on media with PEG for 4 weeks. The maturation media with 87.6 mM sucrose significantly increased the number of ECMs able to form cotyledonary embryos (66.8%) when PEG was included in the maturation medium, but did not affect the average number of somatic embryos. Overall correlation between proliferation diameter during maturation phase and the number of ECMs with somatic embryos (r = −0.40) as well as all significant correlations within individual experimental variants were consistently negative after 8 weeks of maturation, meaning that proliferation growth hampers the formation of somatic embryos. The highest germination percentages 21.6% and 18.2% were obtained when somatic embryos were maturated on media with or without AC supplement, respectively. Lowest germination percentages, 9.1% and 4.4%, were obtained on maturation media with PEG4 and PEG1.

Comparison of embryo yield and pregnancy rate between in vivo and in vitro methods in the same Nelore ( Bos indicus) donor cows

To investigate why the preferred means to produce bovine embryos in Brazil has changed from in vivo to in vitro, we compared these two approaches in the same Nelore cows ( n = 30) and assessed total embryo production and pregnancy rates. Without a specific schedule, all cows were subjected to ultrasound-guided ovum pick up (OPU)/ in vitro production (IVP) and MOET, with intervals ranging from 15 to 45 d between procedures, respectively. To produce in vivo embryos, cows were superovulated and embryos were recovered nonsurgically from 1 to 3 times (1.4 ± 0.6), whereas OPU/IVP was repeated from 1 to 5 times (3.2 ± 1.2) in each donor cow during a 12-mo interval. Embryos obtained from both methods were transferred to crossbred heifers. On average, 25.6 ± 15.3 immature oocytes were collected per OPU attempt. The average number of embryos produced by OPU/IVP (9.4 ± 5.3) was higher ( P < 0.05) than the MOET method (6.7 ± 3.7). However, pregnancy rates were lower ( P < 0.05) following transfer of IVP (33.5%) versus in vivo-derived embryos (41.5%) embryos. Embryonic losses between Days 30 and 60 and fetal sex ratio were similar ( P > 0.05) between in vivo and in vitro-derived embryos. We concluded that in Nelore cows, with an interval of 15 d between OPU procedures, it was possible to produce more embryos and pregnancies compared to conventional MOET.

High success rates with oocyte vitrification in an infertile population

OBJECTIVE: Both infertile and fertile women could benefit greatly from an effective and reproducible method of oocyte cryopreservation. As a complement to traditional assisted reproductive techniques, oocyte cryopreservation allows for storage of supernumerary gametes for infertile patients, provides an opportunity to store oocytes prior to chemotherapy treatment for women with cancer, and offers a valuable tool to the expanding population of fertile women who elect to delay childbearing. The objective of this study is to evaluate oocyte survival and subsequent pregnancy outcomes following oocyte vitrification and warming in an infertile patient population. DESIGN: Descriptive case series. MATERIALS AND METHODS: Eleven infertile women participated in this study. Following oocyte retrieval, all mature (metaphase II) ooctyes were vitrified using the Cryotop technique and stored for an average of 77 ±7 days. Oocytes were warmed, fertilized by ICSI, and developed in culture. Embryo transfer occurred on either day 3 or day 5. RESULTS: The average age of patients was 32.6 ± 4.1 years, with average infertility duration of 3.4 ± 3.0 years. Primary infertility diagnoses included endometriosis (n=4), male factor (n=2), ovulatory dysfunction (n=2), and unexplained (n=3). Overall survival and fertilization of vitrified oocytes were 73.8% (104/141) and 75.0% (78/104), respectively. The cleavage rate on day 3 was 91.0% (71/78) and the rate of blastocyst formation by day 5 of culture was 62.2% (28/45). One patient had no embryo transfer as no oocytes survived the warming process. The average number of embryos transferred was 2.6 on day 3 and 2.4 on day 5. Seven patients became pregnant with an ongoing pregnancy rate of 70.0% (7/10) per transfer, and an implantation rate of 48.0% (12/25) per embryo transferred. All pregnancies are currently ongoing or delivered. CONCLUSIONS: Vitrification is an effective method of oocyte cryopreservation when applied to a typical infertile population. In addition, vitrification has the potential to tremendously expand the reproductive options for women desiring fertility preservation as well as women with a diagnosis of cancer who face gonadotoxic therapy.

Impact of adding recombinant LH versus hMG to rFSH in controlled ovarian stimulation (COS) in embryo morphology and implantation rate: a prospective randomized study

OBJECTIVE: To determine if there are differences in embrionary morphology and implantatoria ability in COS using rFSH and hMG or rFSH and LHr in long protocols with GnRH analogues. DESIGN: Open label, prospective-randomized study. 83 oocyte donors, were included and were randomized into two groups: Group A (n=57): rFSH (150 UI/day) + rLH (37,5 UI/day). Group B (n=26): rFSH (150 UI/day) + HMG (75 UI/day). Pregnancy and implantation rates were compared by Chi Square test and Fisher's exact test considering p <0.05, statistically significant. MATERIALS AND METHODS: In both groups the suppression was performed with GnRH agonists in a long protocol. HCG was administered when at least three follicles ≥ 18 mm were observed. Oocyte retrieval was performed 36 hours later. All embryos were transferred on day 3 of development. RESULTS: There were no differences in both groups in terms of: age, dose of gonadotropins, days of stimulation, estradiol, progesterone and LH on the day of HCG, average number of mature oocytes obtained, fertilization rate, average total embryos obtained, average number of embryos transferred and pregnancy rate per cycle of donor initiated (p>0,05). Ten patients from group A were cancelled because showed a low response. 3 patients from group B were cancelled by the same reason. (p =0.02 difference statistically significant). The pregnancy rate per embryo transfer was 62.07% in group A and 55.81% in group B: (p = 0.17: The difference is not statistically significant). The implantation rate per embryo transferred in group A was 51.8% and in group B was 42.8%: p = 0.09: The difference is not statistically significant). The abortion rate was 21% in group A and 48% in group B for a confidence interval 95% (0.2% to 53.8%), we found a difference of 27% with p = 0,062: The difference is not statistically significant). The rate of Newborn in Group A was 50% and in group B was 30%(p = 0.17: The difference is not statistically significant). CONCLUSIONS: Although starting from the premise expect to find better results with the protocol A (LHr), the results did not show. 1. - The only statistically significant difference found was in the number of cases cancelled (by low response to ovarian stimulation and or low number of oocytes): An increased number of cases in group A (LH), p = 0.02. 2.- We have noticed that both protocols presented similar results in terms of quality embryo, implantation and pregnancy rates.

Low risk of multiple pregnancies in assisted reproductive technology (ART) for women over age 40

OBJECTIVE: Society for Assisted Reproductive Technology (SART) guidelines limit the number of embryos to be transferred in women younger than 40, with the goal of decreasing the rate of multiple pregnancies. However, there are no recommendations for women over the age of 40. We determined the rate of multiple pregnancies in this cohort of older in vitro fertilization (IVF) patients.DESIGN: Analysis of non-donor IVF/ICSI cycle outcomes at 2 university based fertility centers.MATERIALS AND METHODS: Outcomes for non-donor IVF/ICSI cycles performed in women aged 41 – 44 from 2004 to 2007 were analyzed. Clinical pregnancy rate (CPR), defined as a fetal heart beat detected by ultrasound, spontaneous abortion (SAB), live birth rate (LBR), and multiple pregnancy rates were considered for analysis.RESULTS: 363 cycles in patients aged 41-42 and 192 cycles in patients aged 43-44 were initiated, and outcomes are summarized in the table. While the average number of embryos transferred was 3.6 in each age group, the majority of live births were singleton (82% and 77% for age 41-42 and 43-44 respectively), and the chance of a multiple gestation live birth from cycle initiation was 2.5% and 1.6% for age 41-42 and 43-44 respectively. SAB rate in both age groups was 35%.Tabled 141 - 4243 - 44Cycles363192Transfers235149Cycles with no transfer128 (35.3%)43 (22.4%)No. embryos transferred3.63.6CPR (%)21.210.4SAB (%)35.135.0LBR (%)13.86.8- % singleton8277- % twin1623- % triplet20Singleton LBR / transfer (%)17.46.7Twin LBR / transfer (%)3.42.0Triplet LBR / transfer (%)0.40.0Multiple pregnancy LBR / cycle start (%)2.51.6 Open table in a new tab OBJECTIVE: Society for Assisted Reproductive Technology (SART) guidelines limit the number of embryos to be transferred in women younger than 40, with the goal of decreasing the rate of multiple pregnancies. However, there are no recommendations for women over the age of 40. We determined the rate of multiple pregnancies in this cohort of older in vitro fertilization (IVF) patients. DESIGN: Analysis of non-donor IVF/ICSI cycle outcomes at 2 university based fertility centers. MATERIALS AND METHODS: Outcomes for non-donor IVF/ICSI cycles performed in women aged 41 – 44 from 2004 to 2007 were analyzed. Clinical pregnancy rate (CPR), defined as a fetal heart beat detected by ultrasound, spontaneous abortion (SAB), live birth rate (LBR), and multiple pregnancy rates were considered for analysis. RESULTS: 363 cycles in patients aged 41-42 and 192 cycles in patients aged 43-44 were initiated, and outcomes are summarized in the table. While the average number of embryos transferred was 3.6 in each age group, the majority of live births were singleton (82% and 77% for age 41-42 and 43-44 respectively), and the chance of a multiple gestation live birth from cycle initiation was 2.5% and 1.6% for age 41-42 and 43-44 respectively. SAB rate in both age groups was 35%.

The effect of a rise or fall in the serum estradiol level prior to egg retrieval on outcome of in vitro fertilization-embryo transfer cycles in women with diminished egg reserve

The effect of a rise or fall in the serum estradiol level prior to egg retrieval on outcome of in vitro fertilization-embryo transfer cycles in women with diminished egg reserve

Blastomere number and pregnancy rates in the succeeding in vitro fertilization cycle in women who formed all embryos with ≤5 blastomeres

OBJECTIVE: Studies on single embryo transfer showed a marked decreased pregnancy rate following transfer of day 3 embryos with 4 or 5 blastomeres compared to those with 6-9 blastomeres. The objective of the present study was to determine the likelihood of subsequent IVF cycles showing no embryos having greater than 5 blastomeres. Furthermore the study would determine the relative difference in pregnancy rates and implantation rates when embryos with a maximum of 4 or 5 blastomeres were transferred vs. transfers with at least one embryo with 6 blastomeres. DESIGN: Retrospective review. MATERIALS AND METHODS: A 10 year retrospective review was made to identify women having a transfer with no day 3 embryos having 6 or more blastomeres on day 3. All stimulation protocols were used including minimal stimulation. There were no exclusions for serum FSH levels and some cases included serum FSH levels >100 mIU/mL. Viable pregnancy rates at 8 and 12 weeks were determined as well as live delivered pregnancy rates in subsequent cycles according to the group with less vs. more blastomeres. RESULTS: There were 242 IVF cycles identified where the maximum number of embryos had 5 blastomeres. The average number of embryos transferred was 2.0. The clinical 8 and 12 week viable pregnancy rate were 24.8% (60/242) and 22.3% and the live delivered pregnancy rate was 21.1%. The implantation rate was 15.6%. There were 90 of the 191 women not delivering a live baby who attempted another IVF cycle. The majority (82.2%, 74/90) did attain at least one embryo with 6 blastomeres and they did fare slightly better than those in the first cycle in which there were no embryos with 6 cells with an 8 and 12 week viable live delivered pregnancy rate of 29.7% (22/74), and 28.4% (21/74). Interestingly none of the 16 transfers with 4 or 5 blastomeres achieved pregnancies in cycle 2. Sixty-seven women tried 3 or more IVF cycles and 55 (82.0%) had at least one 6-cell embryo. For cycles 2 and beyond the live delivered pregnancy rate per transfer was 32.4% (42/129) with at least one six cell embryo transferred vs. 7.1% (2/28) with ≤5 cell maximum (p<.01, Fisher's exact test). CONCLUSIONS: The majority of women with embryos with ≤5 blastomeres can do better in subsequent cycles. Those that do attain at least one embryo with 6 blastomeres have a reasonable live delivered pregnancy rate. Those who persist with embryos with only 4 or 5 blastomeres have a very low chance of achieving a successful pregnancy.

High viability of biopsied embryos using a new method of vitrification

OBJECTIVE: Preimplantation Genetic Diagnosis and Screening are used in an effort to decrease trisomic conceptions, decrease spontaneous miscarriage and to detect genetic disease. The impact of embryo biopsy on the ability to cryopreserve surplus embryos in PGS cycles needs to be considered, particularly since early reports described reduced viability. The purpose of this study is to demonstrate the effectiveness of the new S3-vitrification method on blastocysts derived from PGS. DESIGN: Retrospective analysis. MATERIALS AND METHODS: From January 2007 through February 2008, 28 patients had cryo-transfers of normal surplus blastocysts - that were vitrified using S3 vitrification during PGS cycles. Embryos were vitrified using several important modifications of regular vitrification. The new S3 approach uses a DMSO-free environment and standard sealable freezing straws. Pre-vitrification steps with this protocol allow for a three-fold duration of exposure time to cryoprotectants. Post-thaw survival was carefully observed. The average number of embryos transferred was 2.1 (range 1-3). RESULTS: All 28 patients had at least one embryo survive for cryotransfer. A total of 67 blastocysts were thawed and 63 of the these survived (94%). 18 of the patients had a positive pregnancy test and 16 of those had a clinical pregnancy (57%). There was one patient who had a spontaneous miscarriage (6.2%). The Implantation rate was 31.7%. We anticipate from this data that the incidence of birth will be in excess of 45%. CONCLUSIONS: This study clearly demonstrates the viability of blastocysts after biopsy on day three and PGS using S3-vitrification. The notion that such embryos do poorly after cryostorage can not be confirmed. The advantage of the new S3 approach is not only demonstrated in increased viability and pregnancy, but also in the shortened learning curve observed when embryologists unfamiliar with this technique were retrained.

Preimplantation aneuploidy testing for infertile patients of advanced maternal age: a randomized prospective trial

To assess the potential benefit of preimplantation aneuploidy testing on the outcome of in vitro fertilization (IVF) for women of advanced maternal age (AMA). Prospective randomized clinical trial. Private IVF clinic. Sixty-two infertile AMA couples undergoing fertility treatment. Fluorescent in situ hybridization (FISH) for chromosomes X, Y, 13, 15, 16, 17, 18, 21, and 22. Preimplantation aneuploidy testing of biopsied blastomeres on day 3 of development. Fertilization and blastocyst developmental rates were similar for the test and control groups: 80% versus 77.4% and 49% versus 48.2%, respectively. The average number of embryos transferred was comparable at 2.2 for the test group and 2.7 for the control group. Implantation rates were also equivalent across the two groups: 37.3% in the control group versus 36.5% in the test group. Nevertheless, the spontaneous abortion rate was observed to be lower for the test group: 25.9% versus 32.26% in the control group. This resulted in an observed increase in delivery rates for the test group: 78% versus 67.74% in the control group. Preimplantation aneuploidy testing does not appear to statistically significantly improve outcome parameters in infertile AMA patients; however, a trend toward a decrease in the spontaneous abortion rate with a subsequent higher delivery rate was observed.

The effects of competition on assisted reproductive technology outcomes

To evaluate the relationship between competition among fertility clinics and assisted reproductive technology (ART) treatment outcomes, particularly multiple births. Using clinic-level data from 1995 to 2001, we examined the relationship between competition and clinic-level ART outcomes and practice patterns. National database registry. Clinics performing ART. The number of clinics within a 20-mile (32.19-km) radius of a given clinic. Clinic-level births, singleton births, and multiple births per ART cycle; multiple births per ART birth; average number of embryos transferred per cycle; and the proportion of cycles for women under age 35 years. The number of competing clinics is not strongly associated with ART birth and multiple birth rates. Relative to clinics with no competitors, the rate of multiple births per cycle is lower (-0.03 percentage points) only for clinics with more than 15 competitors. Embryo transfer practices are not statistically significantly associated with the number of competitors. Clinic-level competition is strongly associated with patient mix. The proportion of cycles for patients under 35 years old is 6.4 percentage points lower for clinics with more than 15 competitors than for those with no competitors. Competition among fertility clinics does not appear to increase rates of multiple births from ART by promoting more aggressive embryo transfer decisions.

Comparison of Blastocyst Transfer With and Without Preimplantation Genetic Screening in Both IVF Patients and Oocyte Donors

Objective: To determine the effect of preimplantation genetic screening (PGS) on pregnancy rate as it relates to blastocyst transfer outcome. Design: Pregnancy rate outcomes were compared for patients and donor cycles having either day 5 blastocyst transfer or day 5 blastocyst transfer with preimplantaion genetic screening. Materials and Methods: We reviewed the records of 561 patients who underwent an IVF cycle with a blastocyst transfer with and without PGS from January 2005 through October 2007 at the Huntington Reproductive Center in Laguna Hills, CA. Group A was composed of the patients who underwent IVF with their own eggs. Group B was composed of the patients who underwent IVF with donor oocytes. Insemination was carried out by ICSI. Fertilized oocytes were cultured in groups in microdrops under oil overlay. Embryos were cultured in Sage cleavage medium and Blast medium (Cooper Surgical, Trumbull, CT) before transfer. All culture was performed at 37°C with 5% CO2 and air. Embryo biopsy was performed in Ca2+/Mg2+-free medium on all embryos with five cells or greater on day 3. Biopsied blastomeres were subjected to fluorescent in situ hybridization (FISH) for chromosomes 13, 18, 21, X, and Y. Embryos were then cultured to the blastocyst stage. All transfers were performed under standard conditions using ultrasound guidance. Average number of mature oocytes, average number of embryos transfered, the percentage of patients with surplus frozen blastocyst remaining after the transfer, and the pregnancy rate were compared. Results: Tabled 1Years 2005–2007Group A, no. of cycles with IVF patientsMean ageAverage no. of mature oocytes% of cycles left with frozen blastocystAverage no. of embryos transfered% of patients with no transferClinical pregnancy rateAverage for day 5 transfer with PGS28834.612.220.8%2.0413.8%46%Average for day 5 transfer without PGS19433.313.960%2.50.005%72.5% Open table in a new tab Tabled 1Years 2005–2007Group B, no. of donor cyclesAverage no. of mature oocytes% of cycles left with frozen blastocystAverage no. of embryos transfered% of patients with no transferClinical pregnancy rateAverage for day 5 transfer with PGS3515.549%2.340%66%Average for day 5 transfer without PGS4418.586%2.040%75% Open table in a new tab Conclusions: In group A, there was a significant difference in clinical pregnancy rate (χ2 = 32.11; P<.0001), in proportion of patients with no transfer (χ2 = 24.95; P<.0001), and in proportion of cycle left with surplus frozen blastocysts (χ2 = 76.05; P<.0001) when comparing day 5 transfer with and without PGS. In group B, there was no significant difference in clinical pregnancy rate (χ2 = 0.43; P=.51); however, there was a significant difference in proportion of cycle left with surplus frozen blastocysts (χ2 = 11.44; P=.0007 [P<.001]). Blastocyst transfer is an effective means to increase pregnancy rate in both IVF patients and in oocyte donor cycles.

Study of Endometrial Receptivity during Implantation in Implantation Dysfunction Mouse

Objective To establish the mice model of implantation dysfunction and to study the endometrial receptivity during implantation in implantation dysfunction mouse. Methods Sexually mature female virgin, Kunming mice were randomly assigned to the control group and the model group postcoitally. The model mice at 9 : 00 AM on d 4 of pregnancy(d 4) were injected subcutaneously with mifepristone. All animals were sacrificed at 9 : 00 PM on d 4 and their uterine horns were examined for the presence of implanted embryos. Histopathology of uterine endometrium was observed by light-microscope. The endometrial expressions of estrogen receptor (ER) and progesterone receptor (PR) assessed by immunnohistochemical SP method. The endometrial expressions of ER mRNA and PR mRNA were assessed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results Compared with control group, implantation rates and average embryo number significently decreased in model group, the development of endometrium was inhibited. In model group, absorbency and area rate of ER and PR in the gland and stroma were lower than those in control group (P Conclusion The endometrial receptivity and implantation decreased in mifepristone- induced implantation dysfunction mouse.

Should embryos destined for cryopreservation be grown to blastocyst?

The favorable pregnancy rates associated with blast transfer were previously outweighed by the difficulty in maintaining embryo development to the blastocyst stage. With the introduction of sequential media, cryopreservation at the blastocyst stage is now more feasible. Our goal is to examine whether there is a difference in the average number of embryos frozen per cycle on day 3 vs. those grown to blast, as well as to assess the difference in thaw survival and clinical pregnancy rates between day 3 embryos vs. blastocysts. Retrospective analysis. All IVF cycles at the Boston IVF center resulting in at least one cryopreserved embryo from Jan 2000 through Jan 2007 were examined to determine the average number of embryos cryopreserved per cycle in the day 3 embryo group and the blastocyst group. Every thaw cycle performed within the same time frame was assessed to determine the average number of embryos thawed, the average number of embryos to survive thaw, and the average percent survival between the day 3 group and the blastocyst group. An average of 3.70 embryos were frozen per cycle in the group undergoing cyropreservation on day 3 vs. 2.67 embryos per cycle among the blastocyst group (P<.05). Average thaw survival among day 3 embryos was 74% compared to 85% among the blastocysts (P=0.05). The clinical pregnancy rate per thaw cycle among the day 3 embryos and the blastocysts was 34.3% and 40.6%, respectively (P=0.36). Table 1Cryopreservation of Embryos Day 3 vs. BlastAve ageTotal # cyclesAve # eggsAve # matureAve # fertilizedAve # embryo transferAve # frozenDay 334.3333213.9111.699.292.333.70Blast33.925016.5614.3011.721.962.67 Open table in a new tab Table 2Thaw Cycle Outcomes Day 3 vs. BlastAve ageTotal # cyclesAve # thawAve # survivedAve # embryo transferAve % survivalClinical pregnancy rateDay 336.426263.472.442.1974%34.3%Blast35.61872.962.211.7785%40.6% Open table in a new tab Although one additional embryo will be gained by performing cryopreservation on day 3 vs. at blast, this appears to be clinically insignificant given that blastocysts are superior in surviving the thaw process as well as in establishing clinical pregnancies. The long time period over which this data has been accrued underestimates, if anything, the true superiority of blastocyst freezing given recent technological improvements in freeze/thaw processes. Routinely growing embryos destined for cryopreservation to blast may result in improved pregnancy rates among patients undergoing thaw cycles.

Outcome of assisted reproduction technology (ART) in women 40 years and older who decide to use their own gametes

For many older women, the option of oocytes donation is not acceptable. We report the outcome with patients older than 40 years attempting ART with autologous oocytes. Retrospective analysis. All couples undergoing ART (IVF or ICSI) between 2004 and 2006, where the female partner was at least 40 years old at the initiation of treatment, were included in this study (n = 520). All patients had a normal serum FSH and estradiol level on day 3. Couples were grouped according to female age (“AGE”). Statistical analysis was done using an One way ANOVA with a Tuckey test a posteriori or Fisher's exact test as appropriate. A total of 396 cycles reached embryo transfer. One hundred and twenty four cycles (23.8%) were cancelled and did not reach embryo transfer due to either the absence of oocytes during follicular aspiration (69.4%, 86/124) or the lack of transferable embryos (30.6%, 38/124). Data regarding average number of oocytes aspirated (“NOA”), percent fertilization rate (“FR”), average embryo quality (“EQ”) (Bolton et al., 1989), average number of embryos transferred (“NET”), percent clinical pregnancy rate (“PR”), percent miscarriage rate (“MR”) and percent birth rate (“BR”) per transfer for all the age and ART groups are shown in Table 1. Tabled 1TableAGENOAFREQNETPRMRBR40 (138)5.3 ± 4.0a78.5 ± 27.015.1 ± 6.72.9 ± 1.728.8a (38/132)0.0 (0/2)15.2 (20/132)41 (94)4.4 ± 2.9a71.4 ± 32.415.3 ± 8.02.7 ± 1.422.3a (21/94)42.9 (9/21)12.8 (12/94)42 (72)4.2 ± 3.2a80.2 ± 30.014.1 ± 6.62.6 ± 4.012.5b (9/72)22.2 (2/9)9.7 (7/72)43 (41)3.6 ± 2.4a78.0 ± 27.016.2 ± 9.12.4 ± 1.59.8b (4/41)50.0 (2/4)4.9 (2/41)44 (27)3.1 ± 1.9b80.8 ± 32.516.5 ± 6.62.4 ± 1.411.1b (3/27)33.3 (1/3)7.4 (2/27)45 (23)3.2 ± 2.5a75.9 ± 28.517.4 ± 8.12.1 ± 1.38.6b (2/23)50.0 (1/2)4.3 (1/23)46 (5)4.1 ± 3.8a66.7 ± 38.014.0 ± 5.83.0 ± 1.90.0 (0/5)–0.0 (0/5)47 (2)3.5 ± 0.7a50.0 ± 0.016.8 ± 6.01.5 ± 0.70.0 (0/2)–0.0 (0/2)Columns with different letters differ significantly (P<0.05). Number of patients between brackets in AGE column. Open table in a new tab Columns with different letters differ significantly (P<0.05). Number of patients between brackets in AGE column. We have achieved an overall live birth rate of 11.3% per embryo transfer (44/389) in patients between 40 and 45 years old, with no births in patients older than 45 years of age (n = 7). Our results confirm the low reproductive success in ART in this large poor prognosis group and the need and value of the oocyte donation option.

Cumulus cell co-culture in the treatment of IVF patients with poor prognosis due to advanced maternal age or two or more failed previous cycles

To determine if cumulus cell co-culture improves clinical pregnancy and implantation rates in IVF patients presenting with advanced maternal age or having two or more prior failed cycles. Retrospective comparative study. This pilot study (January 1, 2007–March 17, 2007) included 21 IVF cycles with patient age ≥38 or with ≥2 previous failed cycles, utilizing Cumulus cell co-culture. During this time period, all patients meeting inclusion criteria were included in the study. The study took place at a private IVF practice. The control group included patient's ≥38 years or with ≥2 previous failed IVF cycles without co-culture from the prior year. Of the 21 cycles; ≥41 yo. (9); 39 yo. (4); 38 yo. (4); 37 yo. (1); 36 yo. (1); 34 yo. (1); 32 yo. (1). Cumulus cells were aspirated at retrieval, trimmed, washed and incubated for 2–4 hours. Just prior to insemination, cumulus cells were separated added to a new dish and incubated overnight at 37°C and 6% CO2. Culture media was removed on Day1; cells were washed and replenished with fresh media. A photograph was taken of the cumulus cells and they were evaluated for attachment. All 2pn zygotes were placed in co-culture. Routine culture procedures were followed from Day 2 to the day of embryo transfer. The difference observed between groups 1 and 2 was not significant with respect to Average patient age (39.4 vs. 40.1); Average number of embryos transferred (3.1 vs. 3.0); Ongoing pregnancy rate (47.6 vs. 43.8); and Implantation rate (26.6 vs. 21.1). The difference in Clinical pregnancy rate (71.4 vs. 50.0) was statistically significant P<0.05. TableGroup 1; Co-Culture (n = 21), Group 2 Control (n = 48)Group 1Group 2Stat. SignificanceAve. Patient Age (yrs.)39.440.1NSAve. Number Embryos Transferred3.13.0NSClin Preg Rate/Tx (%)71.450.0P<0.05Ongoing Preg Rate/Tx (%)47.643.8NSImplantation Rate (%)26.621.1NS Open table in a new tab Cumulus Cell Co-Culture improved the clinical pregnancy rate in IVF patients with poor prognosis due to advanced maternal age or having two or more prior failed cycles.