REPRODUCTIVE BIOLOGY - RESEARCH

O-203 Tuesday, October 21, 2014 04:15 PM

EXPLORING NEW MORPHOKINETIC VARIABLES OF IN VITROAND IN VIVO EMBRYO DEVELOPMENT. M. J. Escriba, L. Escrich,N. Grau, Y. Galiana, Y. Motato, M. Meseguer. Instituto Universitario IVI,Valencia, Spain.

OBJECTIVE: To assess the duration of the second and third cell cycles(cc) as predictors of blastocyst formation and implantation (I).

DESIGN: Retrospective cohort study.MATERIALS AND METHODS: Monitoring of 6970 embryos revealed

that 3107developed into blastocysts (B) and 3863 did not (NB). After trans-fer, 328 embryos were designated as 100%, when the number of gestationalsacs matches with the transferred B or, 0%, when no chemical gestationwas achieved. Timing of cleavage (hrs post-ICSI) from 2 to 8 cells (t2,t3, t4, t5, t6, t7, t8) was determined by time-lapse imaging. The followingvariables were studied: cc2a: cc length of the 1st blastomere to cleavefrom 2 to 3 cells; cc2b: cc length of the 2nd blastomere to cleave from2 to 4 cells; cc3a: b, c, d: 3rd cc duration of the blastomere that cleavesto 5, 6, 7 and 8 cells; acc3: average cc3 (cc3a+cc3b+cc3c+cc3d)/4; blasto-mere synchrony in cleavage at cc2 (SR2¼ cc2a/cc2b) and cc3 (SR3¼acc3/(t8-t3)).

RESULTS: Mean � SD and 95% confidence interval (CI) of variables ac-cording to blastocyst (B or NB) and implantation (0 or 100) status. When sig-nificance between categories was detected, variables were studied byquartiles and the optimal range (OR) for B and I was defined when appro-priate.

Results

Mean � SDaccording to B (95CI) In/out of OR (%B)

ODSRatio OR

Mean� SDaccording to I (95CI) In/out of OR (%I)

ODSRatio OR

Cc2a NB: 9.8�0.1 (9.6-10.0)/B:10.5�3.9 (10.4-10.7)

9.76-12.7 (54.9)/Out (32.6%) 1.6 0: 10.9�3.6 (10.5-11.2)/100:10.5�3.5 (10.2-10.9)

Cc2b NB: 13.1�6.6 (12.9-13.3)/B:12.3�3.9 (12.2-12.4)

11.3-14.3 (51.6)/Out (36.1%) 1.4 0: 12.4�3.6 (12.1-12.7)/100:12.0�3.3 (11.7-12.3)

RS2 NB: 0.7�0.3 (0.7-0.8)/B:0.9�0.2 (0.8-0.9)

%80.5 (28.7)/>80.5 (49.7) 0.8 0: 0.9�0.2 (0.8-0.9)/100:0.9�0.2 (0.8-0.9)

Cc3a NB: 13.4�8.2 (13.1-13.7)/B:13.6�5.6 (13.4-13.8)

11.5-16.5 (55.5)/Out (39.8) 1.3 0: 13.9�4.8 (13.4-14.3)/100:13.6�4.3 (13.1-14.0)

Cc3b NB:17.7�8.4 (17.4-18.0)/B:16.5�5.8 (16.3-16.7)

0: 16.3�4.8 (15.8-16.7)/100:15.6�4.6 (15.2-16.1)

Cc3c NB: 19.3�9.2 (18.9-19.6)/B:17.8�6.7 (17.6-18.0)

0: 17.9�6.0 (17.3-18.4)/100:16.5�4.9 (16.0-17.0)

<21.0 (48.0)/Out (34.1) 1.7

Cc3d NB: 23.5�10.6 (23.0-23.9)/B:21.8�8.9 (21.5-22.2)

0: 21.6�7.9 (20.8-22.4)/100:19.9�7.3 (19.1-20.7)

%15.7 (54.4)/Out (43.2) 1.4

acc3 NB: 17.6�6.5 (17.3-17.8)/B:17.1�5.2 (16.9-17.3)

0: 17.2�4.8 (16.7-17.7)/100:16.1�4.4 (15.6-16.6)

<18.9 (48.5)/Out (38.1)

RS3 NB: 0.7�0.2 (0.7-0.7)/B:0.7�0.1 (0.7-0.8)

%0.7 (47.6)/>0.7 (63.5) 1.5 0: 0.7�0.1 (0.7-0.9)/100:0.8�0.1 (0.7-0.8)

CONCLUSION: cc2 and cc3 are associated with development to B and I,respectively.

O-204 Tuesday, October 21, 2014 04:30 PM

RELATION OF SPERM METHYLATION ABNORMALITY TOMISCARRIAGE VILLUSMETHYLATION ABNORMALITY. A. Sato,a

E. Otsu,a T. Arima,b T. Utsunomiya.a aSt.Luke Clinic, Oita City, Oita, Japan;bTohoku University Graduate School ofMedicine, Sendai City, Miyagi, Japan.

OBJECTIVE: Genomic imprinting, which describes the allele-spe-cific expression of certain genes, accounts for the requirement ofboth maternal and paternal genomes in normal development. Theimprints that are initiated in the germ line persist through preimplan-tation development and involve the formation of an epigenetic markat specific loci in a parent-of-origin-specific manner. DNA methylationis the most well studied epigenetic mark that distinguishes thematernal and paternal alleles of imprinted genes. In this study, we

e70 ASRM Abstracts

examined the methylation profile of four autosomal imprintedgenes in DNA obtained from ART conceptions and matchedparental sperm in order to determine whether these alterations were in-herited.DESIGN: Laboratory experimental study.MATERIALS ANDMETHODS: Between January 2011 and September

2013, 238 cases which resulted in miscarriage following ART wereinvestigated. Of the 238 cases, 48 villus were found to have normal kar-yotype and their origin of sperm. We examined DNA methylation at im-printed gene (H19, GTL2, PEG1 and LIT1) by bisulphate PCRmethylation analysis. Our project was carried out after receiving thepatients’ consent and with the approval of the institutional ethicscommittee.RESULTS: We found 10/48(20.8%) samples that showed abnormal

DNA methylation at one or more imprinted loci. In seven out of theten cases where there were abnormal DNA methylation in the ARTsamples, identical alterations were present in the parental sperm. Sixsamples showed abnormality of the H19 and one sample showed aGTL2 abnormality. All abnormalities were due to paternal imprinttype genes.CONCLUSION: In our previous study of the methylation abnormality in

sperm, maternal type imprint genes (83.3%(20/24)) were found to have ahigher frequency of abnormality than paternal type imprint genes(58.3%(14/24)). Villus and sperm abnormality were found in the paternalimprint genes. Though sperm have both paternal and maternal imprint genes,in this study, abnormalities were seen in the paternal imprint genes, it wasleading us to suggest that maternal imprint genes cannot be the cause ofmiscarriage. Of the 48 samples with normal karyotype examined in thisresearch, 38 were found to also have normal methylation. Further study of

the relationship between imprint abnormality and miscarriages should beconducted.

O-205 Tuesday, October 21, 2014 04:45 PM

THE EFFECT OF AGE ON IMPLEMENTING ONLY VITRIFIEDBLASTOCYST TRANSFER (VFBT) CYCLES AND THE USE OFEUPLOID BLASTOCYSTS (BL) TO OPTIMIZE IMPLANTATIONAND SINGLE EMBRYO TRANSFER (SET). J. B. Whitney,R. E. Anderson, S. Zozula, M. C. Schiewe. SCIRS/SCCRM, NewportBeach, CA.

OBJECTIVE: We have previously shown that VFBT cycles comparefavorably to fresh ET. The purpose of this study was to evaluate whethertrophectoderm biopsy-preimplantation genetic screening (PGS) technol-ogy optimizes pregnancy and implantation outcomes to best support SETefforts?DESIGN: Between 2012 and 2013, all VFBT cycles were split in two

groups. Group 1 (n¼122) patients electively chose to vitrify all fresh

Vol. 102, No. 3, Supplement, September 2014