MATERIALS AND METHODS: Murine zygotes (n¼40) were culturedin commercially available single or sequential media for 5 days. Images(200x, 400x) made using inverted phase contrast microscopes with differ-ential contrast on developmental day 3 (d3) to facilitate quantification ofthe number of embryos developing to the 8-cell stage or greater in thetwo media. On d5, images were made and embryos fixed. Comparingand contrasting embryo images from the two types of media facilitatedidentification of an inner cell mass (ICM), nuclear quality assessment,and quantification of the total number of cells per embryo, ICM and tropho-blast. The least square means (LSM) of total nuclei number, ICM nuclearnumber, and presumptive TE nuclear number between the blastocysts wereanalyzed using a general linear mixed model. An approximate t-test wasused to compare differences in nuclei number (SAS 9.22 PROC MIXED,SAS Institute Inc, Cary, NC). Logistic regression analysis (SAS PROC LO-GISTIC) was used to compare the total embryo numbers per stage per dayon d3 and d5. Hatching/hatched to unhatched blastocysts, TE: ICM andabnormal nuclei:normal nuclei were compared through this same analyticalapproach. A P < 0.05 was considered significant. Deubiquitinating en-zymes localizing in the ICM served as a point of reference for morpho-metric analysis.

RESULTS: Equivalent numbers of embryos reached the 8C stage on d3 inboth single and sequential embryo culture media. More d5 embryos reachedthe blastocyst stage, had greater numbers of normal nuclei, hatched and hadsignificantly more trophoblast cells, but not ICM cells, when cultured in thesingle media compared to the sequential media.

CONCLUSION: Single medium yields higher quality blastocyst embryoson d5 than sequential media. This supports the idea that single stage mediumis more beneficial in human assisted reproduction procedures whencompared to sequential media.

P-236 Tuesday, October 21, 2014

FIRST MORPHOKINETIC ANALYSIS OF BLASTOCYST EXPAN-SION IN HUMAN EMBRYOS OF KNOWN POSITIVE IMPLANTA-TION USING AN EMBRYOSCOPE. T. T. F. Huang. Pacific IVFInstitute, Honolulu, HI.

OBJECTIVE: Using an Embryoscope, the objective was to morphokineti-cally describe features of the expanding human blastocyst cavity in embryosyielding sustained ongoing pregnancies.

DESIGN: Retrospective descriptive study in a private practice setting.MATERIALS AND METHODS: Data were obtained from 27 sequential

egg donation cycles using blastocysts selected for transfer using an Embryo-scope. Day 5 embryos were ranked and selected for transfer by the greatestdegree of blastocyst cavity expansion (BE) with otherwise normal cleavage,ICM and TE characteristics. BE was determined from sequential hourly 2Dcross sectional area (CSA) measurements starting from blastocyst formationup until ET. Sustained pregnancy was defined as clinical detection of abeating heart.

RESULTS: Of 5 single blastocyst transfers (SBT), 4 (80%) resulted in asustained pregnancy (IR¼80%). Of 21 double blastocyst transfers (DBT),21 (100%) resulted in a sustained pregnancy with an IR of 80.4%. (Onepatient had a triple BT but did not become pregnant). Of the DBT’s, 12(57.1%) were twins (100% IR) and 9 (42.9%) were singletons (50%IR). The study confirmed a correlation between the time of the start ofblastocyst cavitation (Tsb) and the time of blastocyst formation (Tb)for both the 100% (r2 ¼ 0.830) and 50% (r2 ¼ 0.893) IR groups. Inthe 100% IR group, the average hourly rate of BE increased over consec-utive 3 hour intervals (557.4 u2 /hr; 916.2 u2 /hr; 1276.3 u2/hr), thentapered to 870.4 u2/ hr during the fourth interval (hours 10-12). Thiswas similar in the 50% IR group. Two morphokinetically distinguishableevents characterized BE. The first was a novel pulsatile, oscillatorypattern of accelerations and decelerations with a periodicity of 2-3 hoursresulting in continuous BE. The second was an occasional, acute contrac-tion of the cavity, ranging from 5-50% in CSA, often occurring severaltimes in blastocysts monitored for up to 22 hours. Remarkably, contrac-tions recovered completely within 1-3 hours. Interestingly, the rates ofexpansion and times to reach given CSA milestones at 1, 5, and 10 hoursfrom Tb did not correlate with the initial time of blastocyst formation inthe cohort of 100% IR blastocysts.

CONCLUSION: This reports the first quantitative description of BE in hu-man embryos yielding sustained pregnancies. Expansion typically acceler-ates in an oscillatory manner, with occasional but reversible collapses in

FERTILITY & STERILITY�

the cavity. Results supports the hypothesis that time-lapse technology iden-tifies novel biological information useful in embryo selection.

P-237 Tuesday, October 21, 2014

MICRO-VIBRATION CULTURE OF HUMAN EMBRYOS IM-PROVES PREGNANCYAND IMPLANTATION RATES. I. El-Dana-souri, N. L. Sandi-Monroy, T. Winkle, K. Ott, C. Krebs,D. H. A. Maas, F. Gagsteiger. Kinderwunsch-Zentrum Ulm/ Stuttgart,Ulm, BW, Germany.

OBJECTIVE: Embryos in vivo are not present in a static condition, sincein the fallopian tube they are exposed to continuous movement, compressioncaused by the cilia and peristaltic movements, and shear stress from tubalfluid flow. In this study, we compared the outcome of human oocytes and em-bryos cultured in a micro-vibration culture system for In Vitro Fertilization(IVF) patients in comparison to the traditional static culture environment.DESIGN: Retrospective analysis.MATERIALS AND METHODS: Oocytes and embryos from 1943 pa-

tients enrolled in IVF treatment from January 2010 to February 2013were cultured in a traditional static culture, while 497 patients’ embryosfrom March 2013 to March 2014 were cultured in a micro-vibration cul-ture. In the micro-vibration group, culture dishes were placed on the topof a platform producing a three-dimensional vibration of 56 Hz for 5 sec-onds every 60 minutes. Patients in both groups were compared according totheir age (< 30; 30-35; 36-38 and >39 years old). The outcomes of micro-vibration culturing were compared for fertilization, blastulation, pregnancyand implantation rates. Variables were analyzed by chi-square test andFisher’s exact test.RESULTS: Pregnancy rates significantly increased in the micro-vibra-

tion culture in younger patient groups compared to the static culturegroups (< 30: 59.4% vs. 38.4%, P¼0.0006; 30-35: 50% vs. 36.3%,P<¼0.0005; 36-38: 42.7% vs. 31.2%, P¼0.03) as did the implantationrate (<30: 54.2% vs. 25.5%, P<0.0001; 30-35: 48.1% vs. 30.1%,P<0.0001; 36-38: 40.3% vs. 26.5%, P<0.0001). The number of trans-ferred embryos and day of transfer did not differ between these groups.For patients >39-years-old, micro-vibration culture tended to increaseboth pregnancy and implantation rates over the static culture but didnot reach statistical significance (20.6% vs. 17.2%, P¼0.41 and 21.1%vs. 15.3%, P¼0.08, respectively). Micro-vibration culturing producedno effect on the fertilization rate. In addition, the overall blastulationrate was significantly higher in the micro-vibration culture compared tothe static culture (40.5% vs. 31.2%, P<0.0001).CONCLUSION: These results demonstrate clearly that the three-dimen-

sional micro-vibration culture of oocytes and embryos significantly increasespregnancy and implantation rates. Mechanical vibration of the embryo maymimic the embryo’s in vivo environment and may cause movement of mediaaround the embryo aiding in refreshing the media surrounding the embryosand diffusion of waste material.

P-238 Tuesday, October 21, 2014

UNDISTURBED EMBRYO CULTURE IN A TIME-LAPSE MONI-TORING SYSTEM IMPROVE EMBRYO QUALITY: A PROSPEC-TIVE COHORT STUDY. W. Han, H. L. Wu, H. Ye, N. G. Huang.Chongqing Maternity and Children Health Care Hospi, Chongqing, China.

OBJECTIVE: To quantify the benefit of culturing embryos in a time-lapsemonitoring system.DESIGN: Prospective cohort study.MATERIALS AND METHODS: One hundred and sixty patients (age

% 35 yrs; BMI: 18-25; 5-15 oocytes; pure tubal factor infertility) wereincluded in this prospective cohort study, and were randomly assignedto the control group (80 patients) or the time-lapse(TL) group (80patients)from April 22 to December 25, 2013. Fertilized oocytes in the TL groupwere cultured in microwell culture dishes and monitored on compact dig-ital inverted microscopes (Primo Vision, Vitrolife). Images were capturedevery 5 minutes .In the control group, fertilized oocytes were cultured inconventional dishes (one embryo in each microdroplet). In both groups,embryo were selected for transfer on day 3, by traditional morphologyassessment. The clinical pregnancy confirmed by the presence of

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