Enhancement of mouse embryo development and implantation by a novel prostacyclin-producing co-culture system -- implication to human IVF

infants are reported by state and plurality. The proportion of ART infants among all infants, multiple-birth, low birth weight, and preterm infants was calculated. RESULTS: Among 3,994,670 infants born in 2011 in the U.S. and Puerto Rico, 1.5 % (59,631) were conceived with ART (range: 0.2% in Puerto Rico to 5% in Massachusetts). ART utilization among states ranged from 313 to 7,502 procedures per million women aged 15-44 years, and was higher than the national rate of 2,401 in 11 of 50 states, many of which were located in the northeast. The national eSET rate among women <35 years was 12.2% (range: 1% in Idaho to 53% in Delaware). Approximately 46%, 31%, and 36% of ART infants were multiple-birth, low birth weight, or preterm infants versus 3%, 8%, and 12% of all infants, respectively. The percentage of preterm ART infants varied from 13% among singletons to 62% among twins and 97% among triplets or higher-order multiples; comparable percentages for all infants were 10%, 57%, and 93%, respectively. Nationally, ART contributed to 20% of multiple-birth (range: 5% in Mississippi to 41% in New York State), 6% of low birth weight (range: 1% in Puerto Rico to 15% in Massachusetts), and 5% of preterm (range: 1% in Puerto Rico to 13% in Massachusetts) infants. CONCLUSION: Wide variations among states were observed in the rates of ART births, utilization, eSET rates, and the contribution of ART to multiple-birth, low birth weight, and preterm infants. Twins and higher order infants were approximately five and seven times more likely to be preterm than were singletons for both ART and all infants. Among ART-conceived infants, greater utilization of eSET, where appropriate, could reduce the contribution of ART to multiple births and preterm delivery. P-549 Wednesday, October 22, 2014 EFFECT OF SPERM HEAD FIRST OR SPERM TAIL FIRST INJECTION INTO OOPLASM ON OOCYTE SURVIVAL, FERTILIZATION AND EMBRYO DEVELOPMENT WITH PIEZO-ICSI IN HUMAN OOCYTES. K. Hiraoka,a S. Kitamura,b M. Kuwayama.a aResearch and Development Division, Repro-Support Medical Research Center, Tokyo, Japan; bART Section, Niji Clinic, Tokyo, Japan. OBJECTIVE: In human oocytes ICSI, it is believed that the sperm should be injected from sperm head first into the ooplasm for fertilization. This is because internalization of sperm into the ooplasm is initiated from sperm head in natural fertilization. The ICSI procedure directly injects the sperm into the ooplasm and bypasses the internalization of sperm head into the ooplasm, so the oocytes could be fertilized if the sperm is injected from sperm tail first into the ooplasm. However, there is no report regarding ICSI results and embryo development of sperm tail first into the ooplasm in human oocytes. Therefore, we investigated the effect of sperm head first injection or sperm tail first into the ooplasm on oocyte survival, fertilization, embryo development with Piezo-ICSI in human oocytes. DESIGN: A prospective randomized study. MATERIALS AND METHODS: Between June 2012 and June 2013, a total of 152 patients were randomly assigned to either sperm head first injection group or sperm tail first injection group with Piezo-ICSI (PMM-150HJ, PRIME TECH). Three hundred and forty two matured oocytes (75 patients; mean age 38.3
4.3) were microinjected by sperm head first into ooplasm and 290 mature oocytes (77 patients, mean age 39.5
4.6) were microinjected by sperm tail first into ooplasm. The rates of oocyte survival, fertilization and good quality day-3 embryo were compared. Good quality day-3 embryos were defined as those having regular blastomeres, <20% fragments and those containing at least 7 cells on day 3. Data were analyzed using Fisher’s exact test. RESULTS: The rates of oocyte survival, fertilization, good quality day-3 embryo per fertilized oocytes of sperm head first injection group and sperm tail first injection group were 99% (339/342) and 99% (288/290), 86% (294/ 342) and 90% (262/290), 69% (202/294) and 68% (179/262), respectively. There were no significant differences in the rates of oocyte survival, fertilization and good quality day-3 embryo between the groups. CONCLUSION: Sperm head first or sperm tail first injection into the ooplasm does not affect oocyte survival, fertilization and subsequent embryo development with Piezo-ICSI in human oocytes. P-550 Wednesday, October 22, 2014 ENHANCEMENT OF MOUSE EMBRYO DEVELOPMENT AND IMPLANTATION BY A NOVEL PROSTACYCLIN-PRODUCING COCULTURE SYSTEM – IMPLICATION TO HUMAN IVF. J.-C. Huang,a W.-S. A. Wun,b K.-H. Ruan.c aObstetrics and Gyenecology, Texas Tech University Health Science Center, Lubbock, TX; bFertility Specialists of Houston, Houston, TX; cPharmacology, University of Houston, Houston, TX.

FERTILITY & STERILITYÒ

OBJECTIVE: Iloprost, a stable analog of prostacyclin has been reported to enhance human IVF success (1). Natural prostacyclin has short half life (3-8 minutes), which renderes experiments with naturally occurring prostacyclin extremely difficult. Our objective was to test a novel de-novo prosatacyclinproducing co-culture system and to determine its impact on embryo development and implantation. DESIGN: Prospective animal trial. MATERIALS AND METHODS: (A) Three to five week old female mice of BL6/C3H mixed genetic backgrounds were used to generated 2-cell embryos (5 units PMSG IP injection initially, 48 hours later 5 units of hCG and mate with male; another 48 hours later, collect 2-cell embryos). Embryos were randomly assigned to method contraol (no plasmid trasnfection), experimental control (feeder layer trasfected with vector) and experimental group (feeder layer transfected with plasmid containing COX-2/PGIS fusion nucleotide). The percentages of completely hatched embryos were compared 96 hour later. (B) Two-cell embryos prepared as above were cultured for 48 hours with feeder layer ransfected with plasmid with (experimental) and without (control) COX-2/PGIS fusion nucleotide (2) before transferring to gestational carriers. The number of gestation sacs were compared 72 hours later. Student’s t-test was used for statistical analysis. RESULTS: In embryo hatching experiment, 50 of the 109 method control embryos hatched completely, 51 of the 110 experimental control embryo hatched completely, and 72 of the 108 experimental embryos hatched completely (p¼0.0028, HR: 1.44, 95% CI: 1.13-1.83). In co-culture/embryo transfer experiment, there were 96 and 94 embryos in control and experimental groups, respectively. Experimental group had more late blastocyst (43 vs. 28 p¼ .024) and fewer morular staged embryos (30 vs. 54, p<0.001). After transferring togestational carriers there were significantly more gestation saces in the experimentla group.

Prostacyclin producing feeder layer enhances embryo implantation

No. embryos transferred No. gestation sacs

Control

Experimental

p

96 32

94 50

n.s. 0.008*

* HR 1.60, 95% CI:1.14-2.24 CONCLUSION: De-novo synthesized prostacyclin from feeder layer cells enhances embryo development and implantation. This serves as a model to provide de-novo synthesized prostacyclin in human IVF culture system. P-551 Wednesday, October 22, 2014 THE EFFECT OF EMBRYO TRANSFER DAY OXYTOCIN ANTAGONIST INFUSION ON IVF OUTCOMES: A SYSTEMATIC REVIEW AND META-ANALYSIS. M. S. Kim,a S. K. Kim,a J. R. Lee,a,b B. C. Jee,a,b C. S. Suh,a,b H. Choi,c S. H. Kim.b aObstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Gyeonggi-do, Republic of Korea; bObstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Republic of Korea; cObstetrics and Gynecology, Inje University Sanggyepaik Hospital, Seoul, Republic of Korea. OBJECTIVE: Several studies demonstrated that transferred embryos could be expelled due to uterine peristalsis and uterine contraction on embryo transfer day was inversely related to the pregnancy rates. Recently there have been increasing attempts of application of oxytocin antagonist (OA, Atosiban) to enhance the pregnancy rate, especially in patients with recurrent implantation failure. The objective of this study was to evaluate the effect of embryo transfer day OA supplementation on IVF outcomes through a systematic review and meta-analysis. DESIGN: Systematic review and meta-analysis. MATERIALS AND METHODS: A computerized search was conducted using three online databases (Cochrane Central Register of Controlled Trials (CENTRAL) in the Cochrane Library, Medline, and Embase). Databases were searched to April 2014 without restriction by language or publication status. A combination of the following key words was used in the search: (atosiban or oxytocin antagonist or oxytocin receptor antagonist or oxytocin receptor) and (embryo transfer or IVF or ICSI or implantation). Studies were selected according to predefined inclusion criteria and meta-analyzed using Review Manager 5.2. Only randomized controlled trials (RCTs) were included in this study.

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