Oct 14, 2013 - RATES. FOLLOWING DAY 6 EMBRYO TRANSFER OR FROZEN EMBRYO. TRANSFER (FET) AFTER BLASTOCYST BIOPSY FOR PREIM-.
CONCLUSION: There is a high rate of blastocyst formation after D5 nonblastocyst biopsy. The practice of biopsying all non-arrested D5 embryos provides the opportunity to have genetic information on all embryos eligible for fresh transfer on D6, potentially adding to the pool of embryos available for transfer. Biopsy of non-arrested D5 embryos may also avoid the need to cryopreserve developmentally competent embryos without ploidy information at the time of transfer. O-120 Monday, October 14, 2013 05:15 PM SIMILAR PREGNANCY AND IMPLANTATION RATES FOLLOWING DAY 6 EMBRYO TRANSFER OR FROZEN EMBRYO TRANSFER (FET) AFTER BLASTOCYST BIOPSY FOR PREIMPLANTATION GENETIC SCREENING (PGS). S. H. Anderson,a T. Stankewicz-McKinney,a M. J. Glassner,a K. Hanshew,a K. Ketterson,b S. Munne.b aMain Line Fertility, Bryn Mawr, PA; bReprogenetics, Livingston, NJ. OBJECTIVE: Blastocyst biopsy and comprehensive chromosome analysis have shown to significantly improve implantation and ongoing pregnancy rates in several clinical randomized studies. However, there is debate on whether it is better to replace on day 6 of culture or in a subsequent FET cycle. Usually centers use one or the other approach, and therefore it is difficult to compare both procedures. Here we present data from a single center with experience in both approaches. DESIGN: Single fertility practice observational study. MATERIALS AND METHODS: Some PGS patients transferred fresh blastocysts on day 6, whereas others transferred thawed biopsied blastocysts for FET. There were no differences between groups in maternal age, number of embryos biopsied on day 5, indications, or chromosomally abnormal embryo rates. Day 5 blastocyst biopsies were sent to a reference laboratory for genetic analysis using array comparative genomic hybridization (aCGH). Results were provided in the morning on day 6 with subsequent replacement before noon for the ‘‘day 6 ET’’ group. Cycles in the ‘‘FET’’ group had their embryos vitrified on the day of embryo biopsy (day 5 or day 6) with replacement of euploid embryos in a FET cycle. RESULTS:
# Cycles Mean Age Mean Blasts Biopsied Mean Biopsied Day 5 Blasts Normal Blasts Abnormal Previous Lost Pregnancies Pregnancy Rate Embryos Replaced Implantation Rate Pregnancies Lost Ongoing Pregnancy
Cycles with No Normal Embryos
FET
DAY 6 ET
20 38.1 2.9 1.9 0.0% 100% 75% -
38 35.3 7.7 5.1 54.7% 45.3% 76% 56% 1.6 57% 0% 56%
46 36.1 5.9 4.6 58.5% 41.5% 62% 70% 1.5 54% 6% 65%
CONCLUSION: The present results do not indicate a significant difference in implantation rates between groups. Most patients prefer fresh day 6 ET of chromosomally screened blastocysts instead of waiting for a FET cycle and incurring additional fees. These results indicated that after blastocyst biopsy and PGS, fresh day 6 embryo transfer is a valid and more economical approach than freezing these blastocysts for a subsequent FET cycle. O-121 Monday, October 14, 2013 05:30 PM COMPARISON BETWEEN MICROARRAY COMPARATIVE GENOMIC HYBRIDIZATION (ACGH) AND FLUORESCENCE IN-SITU HYBRIDIZATION (FISH) FOR PREIMPLANTATION GENETIC DIAGNOSIS (PGD) IN TRANSLOCATIONS CARRIERS. C. M. Sueldo,a T. Escudero,b L. Siano,a A. Finn Bartolucci,a S. Munne,b C. Benadiva.a aObstetrics and Gynecology - Division of Reproductive Endocrinology and Infertility, University of Connecticut, Farmington, CT; b Reprogenetics, Livingston, NJ.
FERTILITY & STERILITYÒ
OBJECTIVE: FISH has been used for many years in PGD for translocations, but recently other techniques have been introduced. The aim of this study was to compare PGD outcomes of patients who underwent FISH or aCGH for detection of reciprocal or Robertsonian translocations. DESIGN: Retrospective Analysis. MATERIALS AND METHODS: Using the database of a reference PGD laboratory, 1151 cycles performed for parental translocations were included. 222 cycles were for Robertsonian translocations, 33 analyzed by aCGH (264 embryos) and 189 by FISH (1366 embryos), while 929 cycles were for reciprocal translocations, 187 analyzed by aCGH (1523 embryos) and 742 by FISH (7347 embryos). RESULTS: Within the Robertsonian group, there was no difference between aCGH and FISH in the rate of aneuploid embryos (29%, 72/248 vs 27%, 342/ 1286), normal/ balanced embryos (32%, 80/248 vs 27%, 348/ 1286), unbalanced embryos (34%, 83/248 vs 32%, 407/1286), nor in the number of ‘‘no result’’ embryos (6%, 16/264 vs 6%, 80/1366). However there was a significantly higher rate of ‘‘complex abnormal’’ embryos detected by FISH (31%, 396/1286 vs 6%, 13/248, p< 0.01). Within the reciprocal group, aCGH detected a higher rate of aneuploid embryos vs FISH + 5 chromosome aneuploidy (17%, 240/1415 vs 7%, 207/3025 p< 0.01; n¼ 277 cycles) and a higher rate of ‘‘complex abnormal’’ embryos (5%, 77/1415 vs. 2%, 175/7067 p< 0.01), as well as a higher number of normal embryos (17%, 246/1415 vs 13%, 913/7067 p< 0.01). However, there was more unbalanced embryos with FISH (80%, 5645/7067 vs 60%, 852/1415, p< 0.01), and in addition, there was more ‘‘no result’’ embryos with aCGH (7%, 108/1523 vs 4%, 280/7347, p< 0.01). CONCLUSION: For Robertsonian translocations, both techniques provided similar results. For reciprocal translocations, aCGH was better at detecting aneuploid embryos, while also providing a higher number of normal embryos. Correlation with miscarriage and live birth rates will be necessary to establish if one technique is superior to the other for PGD of translocations.
O-122 Monday, October 14, 2013 05:45 PM THE BLASTOCOEL FLUID AS A SOURCE OF DNA FOR PREIMPLANTATION GENETIC DIAGNOSIS AND SCREENING. M. Poli,a S. Jaroudi,b J. Sarasa,b K. Spath,a T. Child,a D. Wells.a aNuffield Department of Obstetrics and Gynaecology, University of Oxford, Oxford, Oxon, United Kingdom; bReprogenetics UK, Oxford, Oxon, United Kingdom. OBJECTIVE: This study investigated the possibility of using DNA present in the blastocoelic fluid for preimplantation genetic assessment. DESIGN: Prospective study. MATERIALS AND METHODS: The blastocoels of 15 expanded blastocysts were emptied using a microsuction technique. Paired blastocyst and blastocoel fluid samples were placed in individual tubes. 22 samples (11 pairs) were tested using a specially designed DNA fingerprinting protocol. Analysis involved multiplex-PCR amplification of 8 highly polymorphic microsatellite regions on 8 different chromosomes, including the sex chromosomes. The PCR products were subjected to electrophoresis, revealing the genotypes of each locus and allowing fingerprints to be constructed. Additionally, 8 samples (4 pairs) were amplified using Sureplex (a whole genome amplification method), labelled and tested for aneuploidy using array comparative genomic hybridization (aCGH). RESULTS: In all blastocyst samples, 100% of the polymorphic loci investigated were amplified successfully. However, the number of loci successfully amplified from blastocoel samples was insufficient to allow DNA fingerprinting. Thus the origin of blastocoel DNA could not be confirmed. In the aCGH experiments all samples successfully amplified, but only one out of four blastocoel amplification products completely matched the chromosomal status of the embryo, revealing trisomy for one chromosome. CONCLUSION: From these data, we can conclude that the blastocoel contains a variable amount of DNA, likely of embryonic origin, but the amount is insufficient for the reliable analysis of single copy genes and is therefore unsuitable for preimplantation analysis of single gene disorders. However, we show that in some embryos, a whole genome amplification protocol can allow successful chromosomal screening using aCGH. In the future, further research and development may allow this approach to be used for minimally invasive preimplantation genetic screening. Supported by: Institutional.
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